PAKISTAN SUGAR JOURNAL€¦ · [email protected] ABSTRACT Biomass-based cogeneration in the sugar industry is a useful alternative to meet the shortfall of energy by using

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PAKISTAN SUGAR JOURNALThe first and only research journal regularly published since 1985

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PSJ January-March, 2018 Vol. XXXIII, No.01

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PAKISTAN SUGAR JOURNAL

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International Panel of Referees

Dr. P. Jackson: Principal Scientist, CSIRO, Australia

Dr. Jack C. Comstock: Research Leader, ARS USDA, Canal Point Florida, USA

Dr. William Lee Brusquest Director, CTC, Sao Paulo, Republic of Brazil l

Dr. Raul O. Castillo: Director General, Research Station, Ecuador

Dr. Yong-Bao Pan: Research Plant Molecular Geneticist, USDA-ARS, USA

Dr. James Todd: Commercial Breeder,USDA-ARS,USA

Dr. Niranjan Baisakh: Professor, SPESS, LSU USA

Dr. Aruna Wejasuria, Plant Breeder, Sugarcane Research Institute, Sri Lanka

Dr. Ghazanfar Ali Khan, Additional Secretary Task Force Dept. of Agriculture

Dr. Riaz Ahmed, Chairman, Dept. of Agronomy, UAF

Dr. Nazir Javed, Chairman Dept. of plant pathology, UAF

Dr. Asif Tanvir: Professor, Dept. of Agronomy, UAF

Dr. Kashif Riaz, Assistant professor Dept. of plant pathology, UAF

Dr. Naeem Ahmad Gill, Director, Sugarcane Research Institute, Faisalabad

Dr. Muhammad Ijaz Tabassum, Sugarcane Research Institute, Faisalabad


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CONTENTS Page

Eintensification of power generation in sugar mills through modernization 04

RA Chandgude and PG Patil

Effect of different intercrops on yield, quality and economics of sugarcane 12

Dr. Muhammad Yasin, Abdul Khaliq, Dr. Mahmood ul Hassan and Dr. Naeem Ahmad

Detection Of Genetic Variability Among Sugarcane Somaclones Using 17SSR Marker

Rebina Ferdous, Md. Amzad Hossain, Md Abul Kalam Azad, Mst Kamrun Nahar

SUGAR INDUSTRY ABSTRACTS 26

INTERNATIONAL EVENTS CALENDAR 30

STORY OF SWEETS 31i. Motichoor Ladooii. Chum Chum

GUIDELINES FOR AUTHORS 32


EINTENSIFICATION OF POWER GENERATION INSUGAR MILLS THROUGH MODERNIZATION

*RA Chandgude and PG Patil*Sugar Engineering Division, Vasantdada Sugar Institute, Pune, India

[email protected]

ABSTRACT

Biomass-based cogeneration in the sugar industry is a useful alternative to meet the shortfall ofenergy by using sugarcane residue, an environment friendly fuel available with sugarcane millsacross the globe. The energy obtained from biomass is renewable energy or green energy. Thispaper presents data collected from cogeneration plants in Indian sugar mills and its analysis. Theobservations and findings of this study will be helpful in improving technical parameters in othersugar mills. Power generation, captive power consumption and power export to grid per tonne ofcane crushed are the key factors used for comparison. Power export depends on steam flowcycle, energy conservation in terms of process steam and captive/in house power consumption.Power generation and power export to grid increases with the installation of high-pressure boilerswith pressures of 67 bar, 87 bar, 110 bar and 125 bar and temperatures of 520-560°C.Modernization related to cogeneration in existing and new sugar mills can be implemented toreduce the in-house power and process steam consumption. Energy efficiency in cogenerationplants can be achieved by installation of high-efficiency modern equipment, reactive powermanagement, optimum steam pressure and temperatures, and appropriate loading on turbogenerator sets. Hence, economically, bagasse based co-generation is a viable alternative to meetthe ever increasing energy demand in India. By improving machinery performance, powergeneration is increased and in-house power consumption is reduced, which results in higherpower export to grid. Installation of cogeneration plants in sugar mills increases the revenue ofsugar mill and subsequently improves the rural and national economy. The capital investment isabout INR 50 million per megawatt with a payback period of around 5 years.

Key words Biomass, surplus power, cogeneration, high-pressure boilers

INTRODUCTION

India’s significant andcontinued economic growth isplacing a heavy demand onelectrical energy and this hasincreased the shortage ofpower. The averageelectricity shortage in 2015-16 (up to October 2015) is at2.4% of the normal energyrequirement and about 3.4%at peak load. Electricityproduction in India ispredominantly coal based,and this has high pollutionpotential and there are losses

associated with transmissionand distribution. The focushas now shifted todecentralized and renewableenergy generation. The roleof renewable energy shouldbe considered as alternativesourcesto the existing fossilfuel sources to become a keysolution to the energy needsof the country. Biomass isone of the main sources ofrenewable energy andbagasse is an importantbiomass available inabundance in the sugarindustry. The increase in

bagasse based cogenerationby sugar mills has had apositive impact on the ruraleconomy. This paperpresents data collected fromcogeneration plants in Indiansugar mills and its analysis.The observations andfindings of this study will behelpful in improving technicalparameters in other sugarmills.

GREEN POWER FROM SUGARCANEBiomass is a source ofrenewable energy and can be

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easily converted into energy.The major biomass, i.e.sugarcane residue, is anenvironmental friendly fuel(Anthony et al. 1994)available from sugar mills,and in India, the states ofUttar Pradesh, Maharashtra,Karnataka Tamil NaduAndhra Pradesh, Bihar,Gujarat, Punjab and Haryanahave many sugar mills.Bagasse is considered as themain source of fuel insugarcane-based powergeneration. Sugarcane trash,i.e. the dry and green leavesleft in the field at time ofharvesting, can also becollected along with the caneto provide additional 7.5-10%biomass on cane asadditional fuel to boostcogeneration (Morris andWaldheim 2001). Roots ofcane also have significantfibers, if considered for powergeneration; they can add afurther 2-3.5% additional fiberwhen made available.

It is estimated that totalthermal energy available incane is 640-860 kWh/t ofcane. The present powergeneration level is 110-162kWh/t of cane (Reddy 1994).The sale price of electricityranges from INR 4.50 to 6.59kWh of electricity, dependingupon the tariff fixed by therespective State ElectricityRegulatory Commission.

INDIAN COGENERATION POTENTIAL AND STATUSThere were 538 operationalsugar mills in India during theseason 2015-16, with acrushing capacity of 2.3Mt/day with sugar productioncapacity of over 30 Mt peryear. About one-third of these

sugar mills also have by-product units. The averagecrushing capacity of thesesugar mills is 4000 t/day/unit;they are in private, public andcooperative sectors. In thenear future, 161 new sugarmills will be established. Thepotential from these 538operating and 161 proposedsugar mills stands at 16,404MW of installed capacity ofpower generation and 10,846MW exportable surplus powercapacity during the seasonaloperation. Currently,cogeneration has beencommissioned in 276 sugarmills with installedcogeneration capacity of5,424 MW and 3,539 MWexportable capacity duringseasonal operation. There ispotential for cogeneration ofabout 10,979 MW and 7,307MW of exportable surplusduring seasonal operation.

Sate and Central Governments have encouraged this by:A Renewable EnergyCertificate mechanism thatprovides a market-basedinstrument that can be tradedfreely for additional income.A subsidy from the Ministry ofNew and Renewable Energyup to INR 80.0 million perproject. An income-taxsubsidy for 10 years from theState Government ofMaharashtra. An exemptionon cane purchase tax at 3%on cane price for 10 years.Equity participation of 5% ofproject cost by the StateGovernment of Maharashtra.Switchyard and transmissionline subsidies from StateGovernments.

COGENERATION PRINCIPLECogeneration is theinstallation of steam turbinesand high-pressure boilers forefficient surplus powergeneration that is to beexported to the grid. Theexhaust of steam turbines isused as process heat, utilizedin removing water from juicein order to make dry productssuch as sugar and molasses.The energy in the exhauststeam is utilized for theprocess heat requirements.The application, in which fuelmeets the demand of processheat requirement andelectrical power, is known ascogeneration.

The major advantages of simultaneous production ofheat and power energy are;Environment friendly.Reduced dependency onfossil fuel. Lower fueltransportation cost. Lowergestation period. Reduction inelectrical transmission losses.Increase in local employment.Helps bridge the gap betweendemand and supply ofelectricity. Facilitatesimproved viability andprofitability for sugar mills.

COGENERATION ROUTES ADOPTED IN INDIAN SUGAR MILLSIn Indian sugar mills,cogeneration is carried onthrough two mechanisms,back pressure and theextraction-cum-condensingroute.

Back-pressure routeHere, steam enters theturbine chamber at highpressure and expands to lowor medium pressure. While

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generating power, it alsomeets process steamdemand. This is highlyefficient cogeneration routeresulting in maximum savingin bagasse.

There are two types:Straight back pressure: Thisis the most widely used back-pressure type turbine. Its aimis to expand the availablesteam through the turbinestages and use it for process.Uncontrolled extraction backpressure: In an uncontrolledextraction type, a tap isprovided at a predeterminedstage for drawing out thesteam for process heating.The uncontrolled extractionsteam is used for processand low pressure heatingapplications such as a de-aerator.

Extraction-cum-condensingrouteHere, the extraction-cum-condensing turbines are usedas captive generating setswhere there is variation inprocess steam demand aswell as variation in canecrushing. The surplus steamcondenses in the surfacecondenser. The excesssteam passes through thecondenser, resulting in heatloss in the condenser. Thisroute can be operated in-season and off-season.

There are three types:Straight condensing: In astraight condensing typesteam turbine, the heatenergy of the steam iscompletely converted intokinetic (mechanical energy -torque) energy. The

mechanical energy is utilizedto generate power.

The straight condensing typefinds application in industrieswhere power generation isprime objective, e.g. a captivepower plant.

Uncontrolled extractioncondensing: In anuncontrolled extraction type,a tap is provided at apredetermined stage and apartial stream of steam isdrawn out of the turbine. Theuncontrolled extraction ofsteam is deployed for processapplications (with smallerquantity of steam flow and settemperature requirements)and low pressure heatingapplication such as a de-aerator and process. Thisturbine setup is usuallydeployed in independentpower plants, captive powerplants and cogenerationplants.

Controlled extractioncondensing: A control valveintegrated with the steampath is provided at apredetermined stage. Apartial steam flow is drawnout of the turbine at aconstant pressure as per theprocess requirement. Thereare basically two typescontrol valves, i)diaphragm/grid; and ii)passing/throttle valve, thatcontrol the flow of steam at apreset pressureaccommodating the seasonalvariation in turbine load. Insome cases a nozzle/port isprovided at a predeterminedstage and an uncontrolledhigh-pressure steamflow/bleed is drawn out of theturbine. This tap is provided

before the controlledextraction in the steam pathof turbine. Its utility is foundmainly in low-pressureheating requirements like de-aerator and process.

RESULTS FROM COGENERATION PLANTS

Vasantdada Sugar Instituteconducted a critical analysisof the performance of 34cogeneration projects inMaharashtra during the 2012-13 and 2013-14 seasons. Webased this on data from thegovernment-approved energyauditing agencies. Based on these results, somesugar mills were selected forfurther present study. Theconfiguration and workingresults of these sugar millsare given in Figure 1 andTables 1-3, and calculationsbased on standardassumptions are given inTable 4; the study assumes a3500 t/d basis sugar plantoperating on a 22-h basis,different boiler pressures, millwet bagasse having GCV 9.5MJ/kg, steam 40% on cane,bagasse 28% on cane andturbine extraction-cum-condensing route.

Observations based onthese data are:

Power generation and powerexport are directlyproportionate to pressure andtemperature of boilers(Verbanck et al. 2001).

In-house power consumptionincreases with increase inboiler pressure andtemperature. There isincrease in fixed costs with an

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increase in pressure, butvariable costs decreasesimultaneously. Totaloperating costs decrease withincreases in boiler pressureand temperature.

Revenue generation in a 87bar boiler is INR 189.94 per tof cane - revenue generationis 13.25% higher for a 110bar boiler and 28.12 % higherfor a 125 bar boiler.

PERFORMANCE IMPROVEMENTS IN COGENERATION PLANTS

The key areas that havemajor impact on powergeneration and export are:Use of an efficientcogeneration route.Reductionof process heat energy.Reduction of internal use ofelectrical energy in sugarprocess and cogeneration.Heat recovery from all wastesteam for process andcogeneration.

As the sugar industry is ahighly energy-intensive sectorand consumes energy in allits forms, viz. heat,mechanical and electrical,there is considerable loss ofenergy during the process ofconversion from one form tothe other and the correctselection of the system,technology and equipmentplay a vital role in the overallenergy conservation.

Adoption of modern concepts,such as high pressure boilers,TG sets, condensing andcooling systems, automation,VFD drives, energy efficientmotors, reactive powermanagement, choice of

transmission gears, high-efficiency pumps, correctsizing of all equipment,reducing energy losses dueto leakages, radiation, frictionand reduction in downtimeetc., are the importantaspects to be taken care ofwhile aiming at energyconservation.

By implementing abovemeasures, there will beincrease in overall efficiencyof sugar mill as well ascogeneration plant, mainlyresulting into increase inpower generation and exportper tonne of cane.

ENERGY CONSERVATIONMEASURES

By installing high-pressureboilers and turbo-generatorsets, the power generation incogeneration can beincreased (Kong Win et al.2001) up to 110-162 kWh/t ofcane. By installing efficientmachinery, captive powerconsumption can be reduced(Mydur 1994) to 20-22 kWh/tof cane and for cogeneration5.5-6% on installed power forthe back-pressure route and8-8.5% on installed power forthe condensing route. Suchphase-wise modernizationand energy conservationmethods brought down thesteam % cane to 32-38%from 50% on cane (Avram-Waganoff 1994).

Electrical energy conservation measures:Installation of AC VFD drivesfor mills in place of steamturbines.

Installation of AC or AC VFDdrives for fiberizer/shredder inplace of steam turbines andslip ring motors. Installation ofAC VFD for ID/FD/SA fans,boiler feed water pumps,transfer pumps, bagasse andcoal feeders, cooling towercirculation pumps and fans.

Installation of AC or AC VFDdrives for cane preparatorydevices, cane handlingsystem, feeder tables, canecarriers, inter rake carriers,fuel and ash handling system,screened juice pumps, rawand sulphur juice pumps,massecuite and magmapumps for B and Ccontinuous pans.

Installation of planetary gearboxes for mills, crystallizers,cane carriers, rake carriers,pug mills, sugar mixers,massecuite pumps, sugarconveying system. Carryingout reactive powermanagement by installingcapacitors/harmonic filters inthe form auto power factorcorrection for power factorimprovement and harmonicseparation.Installation of energy efficient motors.

Thermal energy conservation measures:Configuration of theevaporator to be converted toa quadruple/quintuple effectwith extensive vapourbleeding arrangement(Mason 2001).Raw juice primary heating byvapor line juice heater. Rawjuice secondary juice heatingwith condensate. Sulphitedjuice heating in two stageswith vapors from 2nd and 3rdbody of evaporator set.

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Flash heat recoveryClear juice heating by vapors from first body evaporator set.Utilization of 3rd body vaporsfor A and B pans. Low headbatch pans with multidowntake to improvecirculation ratio at low vapourpressures and temperatureswith mechanical circulators.Utilization of 2nd body vaporsfor boiling C pans. Installationof plate heat exchangers forpreheating the boiler make upwater with 3rd bodycondensate. Continuous pansfor B and C massecuites.Waste heat recovery forsulphur burners. Sugarwashing superheated waterby 1st body condensate.

Schemes that determine overall efficiency of cogeneration and sugar plant are:Fiber % Cane - In India, fiber% cane (Mason 1995) variesbetween 12.5 and 18.0%depending upon the variety ofthe cane, age of the cane,time lag between harvestingand crushing, etc. The canethat has high fiber content willhave a lower sugar contentbut it will generate morepower. Hence, using a fiber-rich variety for the sake ofenhancing the power outputis profitable in case of lowsugar prices.

Exhaust Steam Pressure andTemperature - The processexhaust steam pressure andtemperature (Reddy 1994)varies between 1.5 bar and123°C to 3.0 bar and 130°C.At the time of starting thesugar mills, the exhaustpressure is very low and after40 to 45 days the exhaustpressure is on higher side.

Higher the processes exhaustpressure and temperature,lowers the power generation.Ultimately it affects the powerexport. To improve powerexport efficiency always keepclean heating surfaces ofprocess equipment. Astandby evaporator setshould be considered incogeneration sugar plant toavoid cleaning period.Bagasse Saving - In sugarmills, about 0.75% bagcillo isused for vacuum filter, but byinstalling decanters instead ofvacuum filters, this bagacillocan be saved and used togenerate additional power.

Installation of High PressureHeaters - High pressureheaters in boiler circuit willincrease the boiler feed watertemperature. By keepingcontinuous optimum inletpressure and temperature,minimum blow down, goodquality of feed water, keepinginternal and external surfaceclean, etc will also savebagasse.

Bagasse Drying - Bagassedrying increases calorificvalue (Nanda Kumar et al.2001; Maranhao 1994). Thelast mill bagasse with a 50%moisture content has a netcalorific value of 7.54 MJ/kg,but dried bagasse with 10%moisture content possesscalorific value of 15.7 MJ//kg -drying to 10% moisturecontent almost doubles thenet energy value. Every 1%reduction in moisture ofbagasse increases boilerthermal efficiency around by0.5 %.

Diffuser Technology - Themajor electrical energyconsuming section in sugarmill is the Juice ExtractionPlant. In a standard 3500 t/dmilling plant, the powerconsumption is 2050 kWhwhereas in a 3500 t/d diffuserplant power consumption in1475 kWh. The reduction inpower consumption is about575 kWh.

VFD for Fibrizer andShredder – Conventionally, afibrizer is installed with steamturbines and 11 kV slip ringmotors and rotor resistancestarter. Currently, a fiberizeris installed with cage motorsand VFD drives. The losseson account of slip resistanceand low power factor inconventional methods areeliminated in the VFD drivesystem. In addition, theconventional operating rpm ofa slip ring motor is improvedin VFD system, resulting in animproved preparatory index,mill extraction and reductionin power consumption of themill station.

Juice and Water Flow Meters- By replacing theconventional juice and waterweighing scales with flowmeters, power consumptionof pumps for raw juice andimbibition water can besaved. Planetary Gears - Thetransmission efficiency of theworm and worm-wheel driveis about 50-55% and that ofenclosed worm and worm-wheel gear is about 60%.Installing planetary gear bychanging inefficient gearsystem the energytransmission loss can bereduced.

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CONCLUSIONS

There is growing interest inusing a higher proportion ofsugarcane biomass forrenewable energy generationbecause of the rising prices offossil fuels and concern aboutclimate change. Bagasse-based surplus powergeneration has proved to bean economically viablealternative to meet theexisting shortfall of electricalenergy in India. During thefiscal year 2015-2016, theutility sector had an electricityshortfall of 23.6 million MWe.

Electricity from cogenerationis an important alternative tomeet this shortfall. Thecurrent export of power fromsugar mills in India is 3,539MWe and the total potential is10,846 Mwe. High-pressureboilers show higher efficiencyand revenue generation. It isthus recommended to installboilers with pressure 87-125bar and temperature 520-560°C for cogeneration insugar mills. By improvingmachinery performance,power generation isincreased and captive powerconsumption is reduced

ultimately resulting in morepower export to grid. Hence,sugar mills can increase theirprofitability by installingcogeneration andsubsequently undergoingmodernization.

ACKNOWLEDGEMENT

We thank Director GeneralShri Shivajirao Deshmukh forencouraging the study on‘Intensification of PowerGeneration in Sugar Mills byModernization’.

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Bagasse cost INR 2000/t and Electricity tariff INR 6.59/kWh, 160 days season duration are considered for calculations.Fixed cost includes salary, operation and maintenance, manufacturing/water, chemicals, depreciation, and interest on term loan, insurance, administrative overheads.Variable Cost includes bagasse cost during season and off-season operation.Low pressure below 67 bar pressure.

REFERENCES

Anthony J, S. V. Srinivasan, M. Rajavel, 1994. Modern high pressure boilers for cogeneration insugar industry. ISSCT Combined Factory/Energy workshop, pp. 71-78.

Avram-Waganoff P. 2001. Conceptual design of a white sugar factory offering maximumcogeneration. Proceedings of the International Society of Sugar Cane Technologists 24(1): 268-271.

Kong Win KTKE, L. J. C. Autrey, L. Wong Sak Hoi, 2001. Production of electricity from bagasse inMauritius. Proceedings of the International Society of Sugar Cane Technologists 24(1): 282-287.

Lloyd A, J. Hodgson, 2014. Cogeneration efficiency project at Marian Mill. Proceedings of theAustralian Society of Sugar Cane Technologists 36: 444-453.

Maranhao LEC. 1994. Bagasse drying. ISSCT Combined Factory/Energy workshop, pp. 105-117.

Mason V. 1995. Energy conservation and management in Australian raw sugar factories.Proceedings of the International Society of Sugar Cane Technologists 22(1): LXXI-LXXIX.

Mason V. 2001. Electricity cogeneration in cane sugar factories. The output from an ISSCTWorkshop. Proceedings of the International Society of Sugar Cane Technologists 24(1): 180-186.

Morris M, L. W. Waldheim, 2001. Biomass power generation: sugar cane and trash. Proceedingsof the International Society of Sugar Cane Technologists 24(1): 272-274.

Mydur A. 1994. Energy management in Indian sugar factories. ISSCT Combined Factory/Energyworkshop, pp. 399-405.

Nanda Kumar R, L. Nagesh Kumar, 2001. Efficiency improvement in a cane sugar plant using abagasse dryer. Proceedings of the International Society of Sugar Cane Technologists 24(1): 250-251.

Reddy AVR. 1994. Cogeneration in sugar industry. ISSCT Combined Factory/Energy workshop,pp. 273-280.

Verbanck H, P. McIntyre, J. R. Martinelli, 2001. Trends in cogeneration in the Brazilian sugarindustry. Proceedings of the International Society of Sugar Cane Technologists 24(1): 266-271.

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EFFECT OF DIFFERENT INTERCROPS ON YIELD,QUALITY AND ECONOMICS OF SUGARCANE

*Muhammad Yasin, Abdul Khaliq, Mahmood ul Hassan and Naeem Ahmad*Sugarcane Research Institute, Faisalabad

[email protected]

ABSTRACTSugarcane is of great value in the world as it provides sugar to more than half of global population.Cultivated land in the world is decreasing very rapidly due to urbanization, road construction andland deterioration. While the world population is increasing day by day. This world scenariodemands maximum economic returns per acre to feed world population. Intercropping insugarcane is very important in this regard and may become popular among farmers due to longduration and late return from sugarcane crop. Intercropping in sugarcane has potential to providefarmers with maximum economic returns per acre per annum. In a field experiment, differentintercrops were sown in autumn planted sugarcane at research area of Sugarcane ResearchInstitute, Faisalabad, Pakistan during the crop season 2015-16. The intercropping systemscomprised of SC + lentil, SC + linseed, SC + canola, SC + onion and check with alone SC. Theexperiment was laid out in a randomized complete block design with three replications. The resultsrevealed that SC +lentil gave higher cane (138.44 t ha-1) and sugar (16.56 t ha-1) yield withmaximum economic benefits as compared with others.

Keywords: Sugarcane, Intercropping, Economic Return, Cost Benefit Ratio.Abbreviations: SC = Sugarcane; BCR = Cost Benefit Ratio

INTRODUCTION

Sugarcane is an importantcash-cum-industrial crop ofPakistan. It is the mainsource of revenue in Pakistanafter cotton and rice. It is asource of providing rawmaterial to many alliedindustries and employment(Akbar et al., 2011).Sugarcane contributes 3.4%to the value added productsin agriculture and 0.7% togross domestic production.Currently, the area undersugarcane is 1.21 millionhectares and total productionis 73.6 million tons with anaverage yield of 60.42 tonha−1 (Anonymous, 2016-17).

Sustainable agriculture see-ks, at least in principle, to usenature as the model fordesigning agricultural syste-ms. Since nature consistently

integrates her plants andanimals into a diverselandscape, a major tenet ofsustainable agriculture is tocreate and maintain diversity.Intercropping offers farmersthe opportunity to engagenature’s principle of diversityon their farms. Spatialarrangements of plants,planting rates, and maturitydates must be consideredwhen planning intercrops.Intercrops can be moreproductive than growing purestands (Preston, 2003).Intercropping efficientlymaximizes land andproductivity per unit of areaper season (Oad et al. 2001)and owner can obtain moreadded benefits with lowadded costs. Therefore, it isimportant to investigate theadded benefits ofintercropping througheconomic analysis.

Cane is planted in wider rows,and takes several months tocanopy, during which time thesoil, solar energy and muchof the rainfall between therows goes to waste. Anyinter-row crop must thereforemature and be harvestedwithin 90-120 days before thecane canopies (Rathore etal., 1999).Day-by day, the population israpidly increasing whichdecreasing the area undercrop production. Theprerequisite is to increase theproduction and income perunit area. This may bepossible by intercropping(Rehman et al., 2014). Theconventional method ofplanting cane does not permitthe intercrops to grow welldue to shading andcompetition effect.

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The use of leguminousintercrops can help naturallyto increase the availablenitrogen in the soil, therebyreducing the use of inorganicfertilizers (Tosti and Guiducci,2010).The intercrops werealso used in the South Africansugarcane industry tomanage nematodes on smallscale grower farms (Berry etal., 2009).

Autumn-planted sugarcaneoccupies the land for morethan one year and hence thefarmers have no chance totaken other crop in both thewinter and summer season.The growth rate is very slowduring the winter and earlyspring due to prevalence oflow temperature. This periodcan safely be utilized forraising suitable winterintercrops maturing up to theend of April without doingmuch damage to theassociated cane crop (Dhimaet al., 2007).

Pakistan being thesubtropical country with bestgrowing conditions can easilyexploit the potential ofgrowing more than two cropsin a year throughintercropping. This mayincrease production per unitland area with suitable farmmanagement practices.Consequently the presentstudy was designed toexplore the feasibility andscope of different intercropsin sugarcane and theireconomics and assess theireffects on growth, yield andquality of cane.

MATERIALS AND METHODS

The study was conducted atresearch area of SugarcaneResearch Institute,Faisalabad, Pakistan duringthe crop season 2015-16.The experiment was laid outin a randomized completeblock design with threereplications. The net plot sizewas 9.6 m × 5 m. Thetreatments comprised of SC +Lentil, SC + Linseed, SC +Canola, SC + Onion and SCalone as check. Thesugarcane clone S2008-FD-19 with seed rate of 75,000double budded setts perhectare was planted inSeptember 2015 in 120 cmapart double row strips.Trenches were made with thehelp of tractor drawn ridger.

Lentil, linseed, canola weresown in the month of October2015 and onion nursery wastransplanted in the month ofDecember 2015. Half of therecommended seed rate ofintercrops was used i.e.Lentil 20 kg, Linseed 20 kgand Canola 5 kg per ha.Onion was sown by usingnursery. Two lines of eachintercrop were sown inbetween sugarcane.Intercrops were harvested atmaturity during the month ofApril 2016 while thesugarcane crop washarvested in the month ofDecember 2016. Fertilizerwas applied at the rate of175, 115 and 115 kg NPK perhectare. Sixteen irrigationswere applied at differentintervals according to thecrop need and climate.

Data recordingEmergence and tillers per plotwas counted at 45 days and90 days after planting

respectively. Number ofcanes was counted from thetwo strips in each plot at finalharvest and was converted tocanes per hectare. Crop washarvested at maturity bytaking an area of two stripsfrom each plot and cane yieldper hectare was estimated.

Net return was determined bysubtracting the total cost ofproduction from the grossincome of each treatment(CIMMYT, 1988).

Net income = Gross income –Cost of production

Benefit cost ratio wascalculated by dividing thegross income with the totalcost of production.

BCR = Gross income / Totalcost

Statistical analysis The data collected weresubjected to Fisher’s analysisof variance technique andLSD test at 0.05% was usedto compare the differencesamong treatment means(Steel & Torrie, 1984).

RESULTS AND DISCUSSION

EmergenceIt is explicit from the data thatemergence has nonsignificant results in alltreatments. But the maximumgermination of 49.60 % wasattained in SC+ lentil whichwas followed that of by 49.48% in SC + canola. The lowestemergence of 49.25 % wasattained in sole SC. Theresults are in line with thoseof Sain, et al. (2003) who

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reported no significant affectof sole sugarcane anddifferent intercrops onemergence.

Tillers per plantThe summarizedobservations in Table 1regarding average number oftillers per plant recorded atthe time of harvestingrevealed that significantlymore number of tillers perstool (2.20) was produced inthe plots where lentil wasintercropped in sugarcanecrop which was followed thatof by 2.15 in SC + onion. Thedifference in number of tillersper plant of sugarcane withintercrops showed statisticallynon-significant results. Thedata clearly manifested thatintercrops did show nocompetitive effect onsugarcane but lentil improvemore tillers per plant. Tillersper plant are at par in case ofcanola, onion intercrops andsole sugarcane. The lowestnumbers of tillers per plant(2.02) were attained in SC +linseed. These results are incontradict with the findings ofSain, et al. (2003) whoreported smothering andcompetitive affects ofintercrops on tillers per plantof sugarcane.

Cane count (000 ha-1)Regarding the cane count itwas observed that the highestcane count of 140.33thousand ha-1 was recordedin SC + lentil which was atpar with SC + onion with canecount of 139.07 thousand ha-

1. This may be due to moretillers per plant in lentilintercropping in SC. Thelowest numbers of cane count121.51 thousand ha-1 was

recorded in SC + linseed andthese results are in line withBajwa et al., (1992).

Cane Yield (t ha-1)A perusal of data in Table 1showed that Sole SC anddifferent intercrops in SC hada significant impact on caneyield. SC + lentil produced thehighest cane yield with thequantity of 138. 44 t ha-1

which was followed that of by135.55 t ha-1 in SC + onioncrop system. The significantlyhigher sugarcane yield in SC+ lentil is due to higher canecount per ha and more tillersper plant and availability ofsufficient soil nutrients aslentil is a leguminous andrestorative nature crop. Thelowest crop yield of 117.16 tha-1 was attained whenlinseed was sown insugarcane. These results aresimilar to Sain et al., (2003).

Sugar yield (t ha-1)It is clear from the data inTable 1 that sugar yield wassignificantly affected by all thetreatments. The maximumsugar yield of 16. 56 t ha-1

was recorded where lentil wasintercropped in sugarcanewhich was followed that of by15.61 t ha-1 in sugarcane +onion and 15.46 t ha-1 insugarcane + canola cropsystem. The lowest sugaryield of 14.02 t ha-1 wasattained when linseed wassown in sugarcane. On thebasis of these results, it maybe inferred that lentilintercropping in cane will beeconomical and better for thefarmers to get maximumsugar yield. Our results aresupported the findings ofRehman et al., (2014),Sharma et al., (1993) and

Rana et al., (2006) whoconfirmed that lentil + SCproduced higher number oftillers, millable canes andcane yield which ultimatelyleads to highest sugar yield.

Economic analysis:The economics ofintercropping and solesugarcane was worked out intable 2. The economicbenefits got from differentintercrops planted insugarcane were comparedwith the sole sugarcane. Thedata revealed that higheconomic advantage of Rs.542810/- ha-1 with benefit costratio of 3.51 was recorded inthe treatment where lentil wasintercropped in sugarcane.Lentil is a legume intercropenhances soil fertility throughthe excretion of amino acidsinto the rhizosphere. Thenitrogen fixed by the legumeintercrop may be available tothe associated sugarcane inthe current season itself, assugarcane remains in thefield for over nine monthsafter the harvest of thelegumes. Since considerableaddition of nutrient to soil wasresult in more cane and sugaryield per hectare whichultimately leads to moreeconomic benefits thanothers. The lowest net benefitof Rs. 422275/- ha-1 wasattained in sole SC. Ourresults are in line withRehman et al., (2014).

CONCLUSION

It was concluded from theresults of the study thatsugarcane crop producedhigher cane and sugar yieldof 138.44 t ha-1 and 16.56 tha-1 respectively when

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intercropped with lentil. Itgave high economicadvantage of Rs. 542810/-ha-1with benefit cost ratio of

3.51 than other intercropsystems. Economic analysissuggested that a sugarcane-lentil crop system is more

profitable than solesugarcane and otherintercrops.

Table-1 Yield and quality parameters

Treatments Emergence(%)

Tillers/ plant Cane count (000 ha-1)

Cane yield (t ha-1)

CCS% Sugar Yield (t ha-1)

SC + Lentil 49.60 2.20 a 140.33 a 138.44 a 12.76 a 16.56 a

SC + Linseed 49.45 2.02 c 121.57 e 117.16 e 12.25 b 14.02 d

SC + Canola 49.48 2.09 bc 134.51 c 129.83 c 12.74 a 15.46 b

SC + Onion 49.28 2.15 ab 139.07 b 135.55 b 12.74 a 15.61 b

Sugarcane Sole 49.25 2.08 bc 130.46 d 127.30 d 12.35 b 14.77 c

LSD @ 0.05 NS 0.0832 0.6036 1.1252 0.1509 0.2324

Table-2 Economic analysis

TreatmentsCane yield(t-ha-1)

Inter-cropsyield(kg/ha)

Income sugarcane(Rs.)

IncomeIntercrop(Rs.)

Totalincome(Rs.)

Cost ofproductionsugarcane(Rs.)

Costintercrop(Rs.)

Net income/ha (Rs.)

Cost benefit ratio (BCR)

SC + Lentil 138.44 496.03 622980 74405 697385 150575 4000 542810 3.51

SC + Linseed 117.16 535.72 527220 53572 580792 150575 3250 426967 2.78

SC + Canola 129.83 376.97 584235 20733 604968 150575 3200 451193 2.93

SC + Onion 135.55 1295.33 609975 27202 637177 150575 27500 459102 2.58

Sugarcane Sole 127.3 0 572850 0 572850 150575 0 422275 2.80

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REFERENCES

Akbar, N., Ehsanullah, K. Jabran, and M. A. Ali, 2011. Weed Management Improves Yield andQuality of Direct Seeded Rice. Australian Journal of Crop Science, 5, 688-694.

Anonymous, 2017. Economic survey of Pakistan. 2016-17. Finance division. Government ofPakistan.

Bajwa, A. N., M. S. Nazir and S. Mohsin, 1992. Agronomic studies on some wheat basedintercropping systems. Pakistan J. Agri. Sci., 29: 439–43.

Berry SD, P. Dana, V. W. Spaull and P. Cadet, 2009. Effect of intercropping on nematodes in twosmall-scale sugarcane farming systems in South Africa.Nematropica39: 11-33.

CIMMYT. 1988. From agronomic data to farmers recommendations: An Economics trainingmanual. Completely revised edition. Mexico. D. F.

Dhima K. V., A. S. Lithourgidis, I. B. Vasilakoglou and C. A. Dordas, 2007. Competition indices ofcommon vetch and cereal intercrops in two seeding ratio. Field Crops Res 100: 249- 256.

Oad, F. C., A. A. Lakho, G. N. Sohu, M. A. Samo. F. M. Shaikh and N. L. Oad. 2001. Economicsof intercropping of onion with sugarcane. Pakistan Journal of Biological Sciences vol 4 (3).

Preston, S. 2003. Intercropping Principles and Production Practices. Agronomy Systems GuideATTRA-National Sustainable Agriculture Information Service. 1-12.

Rathore O. P., G. K. Nema and H. D. Verma, 1999. Intercropping of medicinal, pulse and spicecrops in autumn-planted sugarcane in Madhya Pradesh. Ind. J. Agron .44(4): 692-695.

Rana, N. S., S. K. Sanjay, Sain and G. S. Panwar. 2006. Production potential and profitability ofautumn sugarcane-based intercropping systems as influenced by intercrops and row spacing. Ind.J. Agron. 51: 31-33.

Rehman, A., A. Ali, Z. Iqbal, R. Qamar, S. Afghanand A. Majid. 2014. Maximum economic returnthrough intercropping of different crops in September sown Sugarcane. P. Sugar. J. V.29 (2): 7-14.

Sain, S. K., N. S. Rana, S. K. Sinha and V. P. Singh. 2003. Parallel multiple cropping of wheat andwinter season sugarcane. Cooperative Sugar. 34(10): 801-803.

Sharma, R. K., K. S. Bangar, S. R. Sharma, H. B. Gwal and H. D. Verma. 1993. Studies onintercropping of pulses in spring planted sugarcane. Ind. J. Pulses Res. 6: 161-164.

Steel, R. G. D. and J. H. Torrie. 1984. Principles and Procedures of Statistics. McGraw Hill BookCo. Inc. Singapore.

Tosti, G. and M. Guiducci, 2010. Durum wheat-faba bean temporary intercropping: Effects onnitrogen supply and wheat quality. Europ. J.Agron.33: 157-165.

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DETECTION OF GENETIC VARIABILITY AMONGSUGARCANE SOMACLONES USING SSR MARKER

*Rebina Ferdous, **Md. Amzad Hossain, ***Md Abul Kalam Azad, ****Mst Kamrun Nahar*,*** Dept. of Biotech. and Genetic Engineering, Islamic University, Kushtia, Bangladesh

**CSO, Bangladesh Sugarcane Research Institute, Bangladesh****Institute of Nano Electronic Engineering (INEE), University Malaysia Perlis, Malaysia

[email protected]

ABSTRACT

The experiment was conducted for detection of genetic variability among ten sugarcanesomaclones including TC2-10(16), TC3-10(16), TC5-10(16), TC8-10(16), TC10-10(16), TC1-10(20) PEG, TC3-10(20) PEG, TC7-10(20) PEG, TC16-10(20) PEG and TC23-10(20) PEG alongwith their parent varieties (Isd 16 and Isd 20) using two Microsatellite (SSR) markers UGSM 565and UGSM 567. The sizes of amplified bands in ten sugarcane somaclones of two varieties Isd 16and Isd 20 ranged from 80 to 650bp. Two SSR markers amplified a total of 58 bands. SSRmarkers UGSM 565 and UGSM 567 amplified 25 and 33 number of bands, respectively. Thehighest number of bands (2.75) per variety was amplified from UGSM 567 marker and lowestnumber of bands (2.08) per variety were recorded from UGSM 565 marker. Genetic diversity orpolymorphic information content (PIC) for UGSM 565 was 0.64 and for UGSM 567 it was 0.69 witha mean of 0.67 for all loci across ten somaclones and their parent varieties evaluated. The highestlinkage distance 6.0 and lowest linkage distance 1.0 were recorded among ten somaclones andtwo parent varieties. Genetic relationships based on UGSM 565 among ten somaclones along withtheir two parent varieties at the average distance of 4.95 showed two major clusters. Thesomaclones TC23-10(20) PEG and TC10-10(16) were separated from other investigatedsomaclones and varieties. On the other hand, based on UGSM 567 genetic relationships amongten somaclones along with their two parent varieties at the average distance of 5.5 showed twomajor clusters and somaclones TC23-10(20) PEG and TC10-10(16) were separated from othersomaclones and their parent varieries respectively. The results revealed that two SSR markerswere able to detect/identify and classify ten sugarcane somaclones along with their parentvarieties indicating genetic differences among somaclones and their parent varieties.

Keywords: Saccharum officinarum, SSR markers, Genetic variability

INTRODUCTION

Sugarcane (Saccharum spp.)is a genetically complex cropof major economicimportance in tropical andsub-tropical countries (Khanet al , 2004 ). Traditional toolsfor sugarcane breeders toidentify different varieties relyon anatomical andmorphological characters(Skinner, 1972). Themorphological characters like

stalk wax, leaf sheath wax,leaf sheath margin, leafsheath hair (pubescence),dewlap appearance, stalkcolor, auricle size and color,and other distinguishingcharacteristics, are generallyused by sugarcane breedersin variety development(LaBorde, Legendre, Bischoff,Gravois, & Robert, 2008). Fordevelopment of improvedvarieties, genotypic studies ofsugarcane is required. To

ensure correct variety identityand its genetic pedigree, aprocedure for accurateidentification using moleculardata is urgently needed (Y.-B.Pan, Cordeiro, Richards, &Henry, 2003; Y Pan, Miller,Schnell, Richard Jr, & Wei,2003; YB Pan, Scheffler, &Richard Jr, 2007). Rapidadvances in the field ofmolecular biology and itsallied sciences made the useof molecular markers a

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routine practice providingplant breeders a precise toolin analyzing genetic diversityfor plant improvement(Andersen & Lübberstedt,2003). The molecularmarkers are of many typese.g. RFLPs, TRAPs, RAPDs,SNPs, simple sequencerepeats (SSRs) and AFLPs.Simple sequence repeats(SSR) markers revealpolymorphisms due tovariation in lengths ofmicrosatellites at specificindividual loci. Microsatellitesor simple sequence repeats(SSRs) are stretches of DNA,consisting of tandemlyrepeated short units of 1-6base pairs in length. They areubiquitous in eukaryoticgenomes and can beanalyzed through PCR(Polymorphic ChainReactions) technology.

In Bangladesh, germplasmcharacterization is mainlybased on agronomic andmorphological traits. So far,all varieties have beenreleased in country on thebasis of agronomic andmorphological traits.Obtaining accurate estimatesof genetic diversity amonggermplasm resources mightbe helpful in sugarcanebreeding. Knowledge ofgenetic diversity andrelationships among breedinggenome, their polymorphicnature, co-dominance andmaterials might have asignificant impact on cropimprovement. Recently,genetic diversity andmolecular characterizationstudy have been initiatedusing RAPD markers. TheSSR marker is importantparticularly for variety

identification by DNAfingerprinting, varietyselection for breedingpurpose, hybridizationevaluation and conservationof their diverse gene pool.Considering the fact, thisinvestigation has beenconducted for detection ofgenetic variability ofsugarcane somaclones usingmicrosatellite markers withthe objectives of (1) detectionof genetic variability ofsugarcane somaclones alongwith their parent varietiesusing microsatellite markers;(2) DNA fingerprinting ofsugarcane somaclones andtheir parent; and (3)determining genetic diversityand relationship amongsomaclones based on clusteranalysis.

MATERIALS AND METHOD

The experiment was carriedout in the BiotechnologyDivision of BangladeshSugarcane Research Institute(BSRI), Ishurdi, Pabna,Bangladesh. The followingmaterials and methods wereconsidered to conduct theexperiment:

Plant materials and samplecollectionFive somaclones of BSRIreleased sugarcane of varietyIsd 16 [TC2-10(16), TC3-10(16), TC5-10(16), TC8-10(16) and TC10-10(16)] andfive somaclones of variety Isd20 [TC1-10(20)PEG, TC3-10(20)PEG, TC7-10(20)PEG,TC16-10(20)PEG and TC16-10(20)PEG], developed byDMSO (Dimethyl sulfoxide)along with PEG (PolyethyleneGlycol), were used as plant

materials for DNA isolation.The fresh tops from the 8month old field grownsugarcane were collected andouterleaf sheaths wereremoved leaving innerspindle to get meristemcylinder. Then meristemcylinder (spindle base) wascut into small pieces (about1.0cm) with sterile scissorsand the required weight (0.2g) was taken using a finebalance.

DNA isolationA modified method of(Aljanabi et al., 1999 ) wasused to isolate total genomicDNA. Cut pieces of meristemcylinder (about 5mm long with3mm dia) weighing 0.2g weretaken in a small mortar (dia6.5cm and 3.5cm depth) andhomogenized with pestle in800μL of extraction buffer(200mM Tris HCl, pH 8.0;50mM EDTA.H2O, pH 8.0;2.2M NaCl; 2% CTAB; 0.06%Sodium Sulfite) until finelyshredded within 40-50second. The grinded samplewas taken into a 2mLeppendorf tube to which wasadded 150μL of each 5%SDS, 10% PVP, 20% CTAB.It was mixed well andincubated at 65°C in a waterbath for 40 minutes. Duringincubation 3-4 times inversionwas done. After incubationsamples were cooled at roomtemperature and equalvolume of Phenol ChloroformIsoamyl Alcohol (25 : 24 : 1)was added mixed well byinversion and centrifuged at10,000rpm at roomtemperature for 30 minutes.Then aqueous phase (about800μL) was recovered andtransferred to a fresh ice-cold2mL eppendorf tube and an

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equal volume of ice-coldisopropanol was addedfollowed by 150μL of 5MNaCl. The samples wereincubated at -20°C for at least1h and centrifuged at10,000rpm at roomtemperature for 20 minutes.The upper layer of solutionwas discarded carefully byusing an adjustablemicropipette and 70% ice-cold ethanol about 2.5 timesof the solution was added. Itwas centrifuged again at10,000 rpm at roomtemperature for 10 minutes.After pellet formation thesolution was discarded fromthe tube carefully so that theDNA pellet remains constantand undisturbed. Then 70%ethanol was added to theslant of the tube. The ethanolwas discarded from the tubecarefully so that the DNApellet remained constant andundisturbed. DNA pellet wasdried for at least 30 minutesputting the tubes upside downon a filter paper. DNA pelletwas re-suspended in 50μL ofTE buffer (10mM Tris HCl, pH8.0; 1mM EDTA.H2O pH 8.0)and stored at -20°C for lateraluse.

Primers usedTwo selected sugarcanemicrosatellite primers(markers) developed byInternational SugarcaneMicrosatellites Consortium,NSW, Australia were used toamplify Simple SequenceRepeats of genomic DNA insix sugarcane varieties. Theprimers were UGSM 565 andUGSM 567 (Table 1). Theprimers were evaluated onthe basis ofintensity/resolution of bands,repeatability of markers and

consistency within individualand potential to differentiatevarieties (polymorphism).

PCR amplification andelectrophoresisPCR amplification was donein an oil-free thermal cycler(Genius, Techne, CambridgeLimited) following PCRprofile of table-2 94ºC for 3minutes (initial denaturation)followed by 35 cycles of 40seconds dennaturation at94ºC, 30 seconds annealingat 55ºC and elongation orextention at 72ºC for 1minutes. After last cycle, afinal step of 7 minutes at 72ºCwas added to allow thecomplete extention of allamplified fragments. Aftercompletion of cyclingprogram, reactions were heldat 4ºC. PCR reactions wereperformed on each DNAsample in a 10μL reactionmixture containing 1.0μL of10x Ampli Taq polymerasebuffer (PCR buffer), 0.6μL of25mM MgCl2, 1.0μL of2.5mM DNTPs, 2.5μL each ofPrimer Forward and Reversefrom 2.5μM stock, 0.2μL of5U/μl Ampli Taq DNApolymerase (Bangalore GeneiPvt. Ltd., India), 2.0μL of25ng/μL genomic DNA and asuitable amount (0.2μL) ofsterile deionized water. Afteramplification, 2µl loading dyewas added to the PCRamplified product and storedat 4ºC for separation usingAgarose Gel Electrophoresis.In each well, 8.0µl of PCRamplified product of eachDNA sample for each primerswas loaded in 1% agarosegel. Electrophoresis wasperformed at 120V for 1.10hours. The DNA ladder forprimer pairs was run along

the sides of the reactions.After electrophoresis, theDNA bands were observedand the data was analyzed.After electrophoresis, the gelwas taken out carefully fromthe electrophoresis chamberand placed in GelDocumentation System(uvitec DBT-2000LS) forscoring of DNA bands. TheDNA was observed as bandand photographed usingalpha view–Fluor Cham FC2software Gel Documentationsystem.

SSR data analysis

SSR data were analyzed forPercentage of PolymorphicLoci (P), Average Number ofAlleles per Locus (A),Average Number of Allelesper Polymorphic Loci (Ap),Average Number ofGenotypes per Locus (G),and Gene Diversity-Polymorphic InformationContent (PIC). Clusteranalysis and Dendrogramswere constructed followingelectrophoresis, and the sizeof amplification products wasestimated by comparingthemigration of eachamplified fragment with that ofa known size fragments ofmolecular weight marker:80bp DNA ladder. All distinctbands or fragments (SSRmarker) were thereby givenidentification numberaccording to their position ongel and scored visually on thebasis of their presence (1) orabsence (0), separately foreach individual variety andeach primer. The scoresobtained using all primers inSSR analysis were thencombined to create a singledata matrix. Linkage

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distances were computedfrom frequencies ofpolymorphic markers toestimate genetic diversity andrelationship between sixsugarcane varieties using theUnweighted Pair-GroupMethod of Arithmetic Means(UPGMA) (Sneath & Sokal,1973) using computerprogram “Statistica”.

RESULTS AND DISCUSSION

DNA fingerprinting of tensugarcane somaclones of twovarieties was performed. Forthis purpose, two SSRprimers were selected aftertest from initially selected fiveprimers because these twoprimers were able to amplifybands in all used sugarcanesomaclones and parentvarieties. Initially five primerswere selected for theinvestigation developed byInternational SugarcaneMicrosatellite Consortium.Molecular characterizationand genetic diversity analysisof 81 sugarcane varietieswere also studied by Muyco(R.R, 2002) using six SSRprimers. He selected thesesix primers from the list of259 primers developed by theInternational SugarcaneMicrosatellite Consortiumbased on observation. Fromthis primer list and accordingto the selection of Muyco, thetwo primers (UGSM 565 andUGSM 567) were selected forthis investigation.

SSR primers withcorresponding bands scored,their size range, number ofpolymorphic bands,percentage of polymorphicbands and number of bands

per variety in two sugarcanevarieties and their respectivesomaclones are presented intable-2. The size of amplifiedbands in ten sugarcanesomaclones and two parentvarieties ranged from 80 to650. The SSR primer pairUGSM 565 revealed the bandsizes ranging from 80bp to650bp while the primer pairUGSM 567 showed a rangefrom 350bp to 650bp. Thiswas perhaps due to thedifferences of the samplesused in this investigation. Theresult was in conformity withthe findings of Yang et al.(Yang, Maroof, Xu, Zhang, &Biyashev, 1994) who pointedout that the range in allelesizes can be influenced bydifferent samples. The twoSSR primer pairs amplified atotal number of 58 bandsfrom ten sugarcanesomaclones and two parentvarieties of sugarcane.RepresentativeElectrophoregrams accordingto primer pairs UGSM 565and UGSM 567 were shownin Figures 1 and 2,respectively For two primerpairs, the total number ofbands produced varied from25 to 33 (Table 2). The primerpair UGSM 567 amplified thehighest number of bands (33)followed by UGSM 565 (25).Taylor and Cordeiro (Corderioet al., 2000) showed that theSSR primer pairs UGSM 567and UGSM 565 producedidentical fingerprints whichpartially supports the presentfindings. The highest numberof bands (2.75) per varietywas amplified from the primerpair UGSM 567 followed byUGSM 565 (2.08). Due to thepolyploidy nature ofsugarcane, the SSR markers

revealed multiple bands perlocus. At South Africa SugarAssociation ExperimentStation (SASEX), Natal,application of 35 sugarcanemicrosatellites identified from1 to 18 alleles per markeracross four varieties. InMauritius Sugar IndustryResearch Institute (MSIRI),Mauritius, number of allelesgenerated per primer pairranged from 9 to 20 using 5primer pairs on 96 sugarcanecultivars (Jannoo, Forget, &Dookun, 2001). At Center forPlant Conservation Genetics(CPCG), Southern CrossUniversity (SCU) in NSW,Australia 3 to 12 alleles perprimer pair were recordedacross five sugarcanegenotypes using 91 primerpairs (Corderio et al ., 2000 ).These results supportspresent findings. In thisinvestigation, highestpercentage (16%) wasrecorded from the primer pairUGSM 565 which producedthe lowest number of totalbands (25) whereas lowestpercentage (6.06%) ofpolymorphism was recordedfrom the primer pair UGSM567 which produced thehighest number of total bands(33). This variation in bandformation and polymorphismmay be due to variation ofprimer pairs. The two SSRmarkers UGSM 565 andUGSM 567 discriminated17.33% and 22.92% ofvarieties and somaclonesevaluated, respectively. Thisdiscrimination may be due tothe differential response ofthe primer pairs with thesomaclones and parentvarieties.

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Both the two SSR markersgenerated multiple fragmentsamong ten sugarcanesomaclones and two parentvarieties. The respectivegenetic diversity orpolymorphic informationcontent (PIC) per marker ofUGSM 565 and UGSM 567was recorded as 0.64 and0.69 for all loci across the tensomaclones and two parentvarieties evaluated (Table 3).The primer pair UGSM 565showed PIC value 0.64 whilethe marker UGSM 567produced PIC value 0.69. Themost polymorphic SSRmarker was associated withthe highest number ofpolymorphic bands detected.The primer pair UGSM 565was most polymorphic markeracross ten somaclones andtwo sugarcane varieties withPIC value of 0.64. The PICvalues are dependent ongenetic diversity ofsomaclones and varietiesunder study. According toGarland et al, (Garland,Lewin, Abedinia, Henry, &Blakeney, 1999) a highproportion of closely relatedgenotypes would have effectof lowering PIC values. In thisstudy, mean PIC valueamong ten somaclones andtwo varieties was 0.67indicating a high level ofvariability present in varietiesand somaclones based ontwo SSR primers.Comparable results were alsoreported by Corderio et al,(Corderio et al ., 2000 ) insugarcane where they usedthe SSR primers.

Genetic relationships basedon UGSM 565 among tensomaclones along with their

two parent varieties at theaverage distance of 4.95showed two major clusters(C1 and C2) presented inFigure 3. At linkage distanceof 4.00 the cluster C2

produced sub-cluster SC1 andSC2. Finally, sub-cluster SC2

produced sub-sub-clusterSSC1 and SSC2 at the linkagedistance of 2.0. The majorcluster C1 separated thesugarcane somaclone TC23-10(20) PEG from the othersomaclones and the parentvarieries. On the other hand,the major cluster C2 separatedthe somaclone TC10-10(16)from other somaclones andthe parent varieties. Based onUGSM 567, geneticrelationships among the tensomaclones along with theirtwo parent varieties at theaverage distance of 5.5showed two major clusters(C1 and C2) presented inFigure 4. At the linkagedistance of 1.00, the clusterC1 produced sub-cluster SC1

and SC2 and at the linkagedistance 4.80, the C2

produced sub-cluster SC1 andSC2. Finally, sub-cluster SC2

of the cluster C2 producedsub-sub-cluster SSC1 andSSC2 at the linkage distanceof 1.0. The major cluster C1

separated the sugarcanesomaclone TC23-10(20) PEGfrom the other somaclonesand the parent varieries. Onthe other hand, the majorcluster C2 separated thesomaclone TC10-10(16) fromother somaclones and theparent varieties. In bothmarkers, results of DNApolymorphism of the tensomaclones and twosugarcane varieties provethat they bear the genetic

diversity of somaclonesTC23-10(20) PEG and TC10-10(16) from otherinvestigated somaclones andvarieties. Hence, it is clearfrom figure 3 and 4 thatsomaclones TC23-10(20)PEG and TC10-10(16) shownto be outliers in dendrogramand distantly related with restof somaclones based ontheir genetic distances. Theresult was in partialagreement with findings ofShahnawaz (R.M.S, 2006)who analyzed DNApolymorphism of the fourvarieties/accessions andfound that the variety Isd 16was separated from the othervarieties in a major clusterwhile the remaining threewere in another cluster. Healso used variety Isd 20having superior performancesagainst biotic and abioticstresses than the othervarieties. It lied in the sub-cluster SC1 and wasseparated from other varietiesat the linkage distance of11.0.

CONCLUSION

The results of the presentinvestigation revealed thattwo SSR primers were able todetect or identify and classifyten sugarcane somaclonesalong with their parentvarieties indicating geneticdifferences among sugarcanesomaclones and their parentvarieties. Therefore, detectionof genetic variability amongsugarcane somaclonesshould be continued in orderto determine their geneticdistances and relationshipsamong them usingmicrosatellite markers.

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Table-1 Parameters of primers sequence of two sugarcane microsatellite primers from the International Sugarcane Microsatellite Consortium, NSW, Australia

Primer Code Sequence (5/-3/) G+C Content (%)UGSM 565 Forward: 5'-CAT AGC AAG CAC CAC CTC TC-3'

Reverse: 5'-TCT TCT TCT CGT CCA CCC-3'55.0055.56

UGSM 567 Forward: 5'-CTT CAT ACG CCA CCT TCT C-3'Reverse: 5'-CAA ATG TTC ACT CGC ATC A-3'

52.6342.10

Table-2 Microsatellite primers with corresponding bands scored, their size range,number of polymorphic bands, polymorphism and number of band per variety together with variety distinguished in two sugarcane varieties and ten somaclones

Primer Codes

Sizeranges

(bp)

Totalnumber ofband score

Number ofpolymor-

phicbands

Polymor-phism

(%)

Numberof band

pervariety

Varietydistinguished

(%)

UGSM 565 80-650 25 4 16.00 2.08 17.33UGSM 567 350-650 33 2 6.06 2.75 22.92

Total 58 6 - - -

Table-3 Microsatellite primers with Polymorphism Information Content (PIC).

Primer Codes PIC(Polymorphism Information Content)

UGSM 565 0.64UGSM 567 0.69

Figure-1 SSR profile of ten somaclones along with their two parent varieties based on Primer pair UGSM 565

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Fig. 2 SSR profile of ten somaclones along with two parent varieties of sugarcane based on primer pair UGSM 567

Tree Diagram for 12 Variables

Unweighted pair-group average

Squared Euclidean distances

Linkage Distance

TC23-10(20)PEG

TC16-10(20)PEG

TC7-10(20)PEG

TC3-10(20)PEG

TC1-10(20)PEG

Isd 20

TC10-10(16)

TC8-10(16)

TC5-10(16)

TC3-10(16)

TC2-10(16)

Isd 16

0 1 2 3 4 5

Fig.3 Tree Diagram for ten sugarcane somaclones and two varieties based on Unweighted Pair-group average Squared Euclidean distances using primer pair UGSM 565

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Tree Diagram for 12 Variables

Unweighted pair-group average

Squared Euclidean distances

Linkage Distance

TC23-10(20)PEG

TC16-10(20)PEG

TC7-10(20)PEG

TC3-10(20)PEG

TC1-10(20)PEG

Isd 20

TC10-10(16)

TC8-10(16)

TC5-10(16)

TC3-10(16)

TC2-10(16)

Isd 16

0 1 2 3 4 5 6

Fig.4 Tree Diagram for ten sugarcane somaclones and two varieties based on Unweighted Pair-group average Squared Euclidean distances using primer pair UGSM 567

REFERENCES

A, B. (2000). Application of sugarcane microsatellites at SASEX. The Satellite 3, 1, p. 3.

Aljanabi, S., L. Forget, and A. Dookun, (1999). An improved and rapid protocol for the isolation ofpolysaccharide-and polyphenol-free sugarcane DNA. Plant Molecular Biology Reporter, 17(3),281-281.

Andersen, J. R., and T. Lübberstedt, (2003). Functional markers in plants. Trends in plantscience, 8(11), 554-560.

Corderio, G., G. Taylor, and R. Henry, (2000). Characterization of microsatellite markers fromsugarcane (Saccharum sp.), a highly polyploidy species. Plant Sci, 155, 161-168.

Garland, S. H., L. Lewin, M. Abedinia, R. Henry, and A. Blakeney, (1999). The use ofmicrosatellite polymorphisms for the identification of Australian breeding lines of rice (Oryza sativaL.). Euphytica, 108(1), 53-63.

Jannoo, N., L. Forget, and A. Dookun, (2001). Contribution of microsatellites to sugarcanebreeding program in Mauritius. Proe. Int. Soc. Sugarcane Technol, 24, 637-639.

Khan, I., A. Khatri, G. Nizamani, M. Siddiqui, and M. Khanzada, (2004). In-vitro Culture studies insugarcane. Pak J Biotech, 1, 6-10.

LaBorde, C., B. Legendre, K. Bischoff, K. Gravois, and T. Robert, (2008). Sugarcane VarietyIdentification Guide 2008. Louisiana State University AgCenter Publication, Baton Rouge.

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Pan, Y.-B., G. Cordeiro, E. Richards, and R. Henry, (2003). Molecular genotyping of sugarcaneclones with microsatellite DNA markers. Maydica, 48(4), 319-329.

Pan, Y., J. Miller, R. Schnell, E. Richard Jr, and Q. Wei, (2003). Application of microsatellite andRAPD fingerprints in Florida sugarcane variety program. Plant and Animal Genome Abstract XIAbstract Book W, 189, 43.

Pan, Y., B. Scheffler, and E. Richard Jr, (2007). High-throughput molecular genotyping ofcommercial sugarcane clones with microsatellite (SSR) markers. Sugar Tech, 9(2-3), 176-181.

R. M. S, S. (2006). DNA Isolation, Quantification and Fingerprinting Using RAPD Markers ofSugarcane (Saccharum officinarum L.). A thesis of Master’s of Science (M. Sc.) in Biotechnologyand Genetic Engineering Department of Islamic University, Kushtia, Bangladesh.

R. R, M. (2002). Genetic diversity in sugarcane (Saccharum officinarum L.) from the activegermplasm collection of Philsurin based on coefficient of parcentage, morphological traits andDNA microsatellite markers.

Skinner, J. (1972). Description of sugarcane clones. III. Botanical description. Proceedings of theInternational Society of Sugar Cane Technology, 14, 124-127.

Sneath, P. H., and R. R. Sokal, (1973). Numerical taxonomy. The principles and practice ofnumerical classification.

Yang, G., M. S. Maroof, C. Xu, Q. Zhang, and R. Biyashev, (1994). Comparative analysis ofmicrosatellite DNA polymorphism in landraces and cultivars of rice. Molecular and GeneralGenetics MGG, 245(2), 187-194.

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Sugar INDUSTRY ABSTRACTS

Isolation and preliminary biochemical characterization of nitrogen-fixing bacteria belongingto three genera obtained from sugarcane in Colombia

Jennifer Roa, Marcela Cadavid, Fernando Muñoz, Héctor A Chica and Carlos A ÁngelProceedings of the International Society of Sugar Cane Technologists, volume 29, 2016

Sugarcane is an important crop in Colombia, intensively planted to about 228,000 ha along theCauca River Valley. To reach profitable yields, high and in some cases excessive nitrogen (N)fertilizers are applied, increasing production costs and risks for environmental and watercontamination. Looking for strategies to reduce N applications within a sustainable and integratedcrop management, the role of N-fixing bacteria is being explored. The objective of this study wasto isolate and characterize native cultures of three recognized N-fixing bacteria and plant growthpromoter genera such as Azospirillum, Azotobacter and Gluconoacetobater, and to determinetheir association to four of the most important and widely planted CENICAÑA-Colombia (CC)varieties (CC85-92, CC84-75, CC93-4418, and CC01-1940). To cover most environments alongthe valley, 108 fields were selected using geographical information system ArcGIS ®. Individualsamples consisted of 20 plants were selected randomly according to a predetermined method,sampling roots, stems, leaves, and rhizospheric soil from each plant and site. Samples wereprocessed at CENICAÑA’s Plant Pathology Lab, plating on selective culture media LGI-P, Ashby,and NFB, among others for each bacteria genus. Morphological and biochemical characterizationfor α and β-proteobacteria were undertaken by triplicate, including aerobic/microaerophilic,motility-helical-shaped rods and cocci, Gram stain, and oxidative-fermentative metabolism ofdifferent carbohydrates and organic salts. In addition, production of growth-promoting substanceswas also evaluated. Comparisons were undertaken against type cultures of Azotobacterchroococcum NCBIM 8002, Azospirillum brasilense NCBIM 11860, Gluconacetobacterdiazotrophicus NCBIM 12985, among other bacteria. Out of 143 obtained isolates, 111 werecharacterized. Cluster analysis identified three groups that explained 91% of biochemical andmetabolized carbohydrates variation. Quantification of indole-3 acetic acid as growth promotingsubstance showed also high variation with similar contents between Azotobacter spp. andAzospirillum spp. isolates, but significant statistical differences (α=0.05) to Gluconacetobacter spp.with lower production. Sequencing of 16S rDNA to identify species as well as acetylene reductionassays to determine N-fixing performance for these isolates is in progress. Fourteen isolates fromthe three genera were identified as promising candidates for further studies.

Use of apparent soil electrical conductivity to improve sugarcane nutrient management inFlorida

Hardev S Sandhu, Maninder P Singh and James M McCrayProceedings of the International Society of Sugar Cane Technologists, volume 29, 2016

Precision agriculture is considered to be one of the most promising approaches for sustainablefarming, but it requires efficient methods for accurately measuring within-field variations in soilphysical and chemical properties. Apparent soil electrical conductivity (ECa) is a quick indirectmeasurement of soil EC with a sensor (e.g. EM-38), and the latter is used to determine spatialvariations of this parameter in the field without extensive soil sampling. Correlation of ECa valueswith different soil nutrients in organic and mineral soils in southern Florida can help in determiningits potential use to improve nutrient management in sugarcane. Data on ECa and different soil

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variables were collected from several 5-15 ha fallow sugarcane fields in Palm Beach (organic soil,Histosols) and Hendry (mineral soil, Entisols) counties of Florida in 2014 and 2015. Soil sampleswere analyzed for pH, P, Ca, Mg, S and Si in soil testing lab at the Everglades Research andEducation Centre, Belle Glade. In organic soils, the bulked soil data showed significantcorrelations between ECa and each of soil pH (r=0.72), Mg (r=0.62), Si (r=0.52) and Ca (r=0.35).In mineral soils, there were significant correlations between ECa and each of pH (r=0.75),PMehlich-3 (r=0.73), Ca (r=0.58), and Si (r=0.46). Grouping the fields into different zones basedon their location changed the correlations for the tested variables. PC-stepwise regressionanalysis indicated that soil pH was the major contributor to the variability in soil EC a in both soiltypes. Results showed that the correlations between ECa and the measured soil parameters werenot consistent through all the tested zones or the fields. Therefore, ECa may be more preciselyused in management of these parameters at zonal or field level. In bulk soil, the correlationsbetween ECa and soil pH were highest in both soils and that supports the need of furtherexploration of ECa maps in soil pH management across a wide range of sugarcane fields inFlorida. Alternatively, ECa may be used in conjunction with soil sampling to determine the spatialvariability of the soil variables and thereafter be used for precise management of differentnutrients. Further research is needed to determine the relationship between EC a and sugarcaneyield, and for the use of yield maps in ECa map calibrations.

Sugar storage in silos

M Schuermann, R Timmers, P Avram and B MorgenrothProceedings of the International Society of Sugar Cane Technologists, volume 29, 2016

There are large differences in the usual storage of white sugar from beet and cane. Beet whitesugar is usually stored in silos. Cane white sugar is commonly filled in bags and stored inwarehouses. Sugar quality is much better using silo storage instead of warehouses in terms offluidity and aspects of hygiene as well as food production. Silo-handled sugar can either be filled inthe usual packaging machines or be filled and transported in road tankers – the latter is veryeconomical especially for industrial use. In contrast to the storage in warehouses, sugar storage insilos has to fulfill some important parameters. This paper highlights particular items such asprimary screening, temperature and moisture, silo filling procedure, sugar conditioning, dedustingand discharge. The aim of this paper is to provide recommendations for the cane sugar industry toimprove their sugar quality by using silos for storing white sugar.

Overview of crystallization in vacuum pans in the Colombian sugar industry

JD Tascon, JM Latorre, JG Rodriguez and NJ GilProceedings of the International Society of Sugar Cane Technologists, volume 29, 2016

This paper presents an analysis of vacuum pan crystallization in Colombian factories. Attention tothis process is given because sucrose losses in molasses represent up to 50% of the total lossesof sucrose. For this research we gathered several data of boiling stages and strikes in Colombianfactories. A representative boiling scheme for two and three boiling stages is presented withaverage data of brix and purity. Typical conditions of actual equipment in vacuum pans aresummarized. Additionally, we captured process data and physicochemical properties under normalindustrial conditions. Sugar crystallization was evaluated in terms of crystal quality, sucroserecovery and energy consumption. We propose best operational practices that aim at profitableprocessing of sugar even under manual control of vacuum pans. The strategies should lead toachieving higher or constant recovery rates and a reduction in boiling time of up to 15%.

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Crop diversity in sugarcane: effect of mixed cultivars on the growth and yield of sugarcane

H Takaragawa, K Watanabe, J Thanankorn, M Nakabaru and Y KawamitsuProceedings of the International Society of Sugar Cane Technologists, volume 29, 2016

Sugarcane is usually grown as a monoculture over a large area of the world. Monoculture of cropssuch as sugarcane easily destroys the biodiversity of the farmland during cultivation. Forsustainable agriculture, we have to consider the concept of biodiversity in agriculture in terms ofagro-biodiversity, agro-ecosystem and crop diversity. Recently mixing cultivars has beendemonstrated to have potential as a new method to achieve high and stable yield in some crops.Mixing cultivars with different traits of tolerance to stresses or different growth rates createscultural breaks in the field to prevent diseases and harmful insects from spreading and usespositive effects of competition and compensation between cultivars to increase growth and yield.Our team is trying to determine the effects of mixed cultivars on the growth and yield in sugarcane.In experiment 1, two cultivars with different canopy structures were selected, one has high stalkweight, the other has many tillers. These were mixed by plant or row in Okinawa, Japan. Light-extinction coefficient was lower in the mixture-by-row and higher in the mixture-by-plant comparedwith monoculture. Stalk length of one cultivar in the mixture-by-row was shorter, which overallresulted in the better light-intercepting characteristics of this canopy. This was consistent with anincrease in the number of millable canes in the mixture-by-row. A productive structure diagramshowed that the effect of mixed cultivars on canopy formation depended on the way cultivars weremixed. In experiment 2, two high stalk weight cultivars were mixed by plant in Tokunoshima,Japan. The results showed a reduction in the number of stalks in the mixture due to the reducedestablishment of one cultivar. However, the stalk weight in the mixed cane increased and the yieldof the mixture was almost equal to that of monocropped cultivar. Our results indicated that habitatsegregation and a compensation effect may have occurred in the sugarcane canopy of mixedcultivars.

Screening of phosphate-solubilizing bacteria from sugarcane rhizospheric soil and theirabilities to improve growth and yield of sugarcane

C Thongponkaew, N Chittamart, S Tawornpruek and P PinjaiProceedings of the International Society of Sugar Cane Technologists, volume 29, 2016

Phosphorus (P) is one of the three most important mineral nutrients in crop production. Only 0.1%of the total P in soils is available to plants due to its poor solubility and P fixation in soil. Currently,enhancement of microbial activities by biofertilizers in sustainable sugarcane production hasbecome more attractive. Phosphate-solubilizing bacteria (PSB) are interesting microorganismsthat transform insoluble P to plant-available P. This study aimed to isolate and screen for bacteriathat have high phosphate-solubilizing activity from major sugarcane rhizospheric soils in Sa KaeoProvince, Thailand, and to examine the potential of PSB to promote plant growth. We obtained1,242 isolates and 1,156 of these (98% of total isolates) enabled tricalcium phosphate to besolubilized. However, only 151 isolates exhibited high phosphate-solubilizing activity. These 151isolates were then evaluated for nitrogen (N) and carbon (C) source utilization using UPGMAanalysis. The results indicated that the bacterial isolates could be categorized into two clusterswith a maximum similarity value of 55%. The biochemistry patterns showed that the strains couldbe divided into 34 unique groups. All of these isolates were characterized for their N-fixing ability,and indole-acetic acid and gibberellin producing ability. Eleven of these isolates (33%) werecapable of fixing N, and 34 isolates (100%) can produce both IAA and GA. Among these isolates,the five highest phosphate-solubilizing isolates are Tpk3-053, Tpk4-083, Wi1-013, Ch1-021 andMki4-010. They can solubilize phosphate ranging from 447 to 748 μg.mL-1, produce IAA rangingbetween 1.42-11.83 μg.mL-1 and GA at 4.29-4.76 μg.mL-1. Although these five isolates are suitable

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for developing as biofertilizer, the isolates need to be tested for their ability to promote growth insugarcane under greenhouse and field conditions.

Transcriptome analysis of host-pathogen interaction between sugarcane andColletotrichum falcatum by Suppression Subtractive Hybridization and Illumina sequencing

R Viswanathan, M Sathyabhama, P Malathi and A Ramesh SundarProceedings of the International Society of Sugar Cane Technologists, volume 29, 2016

A high throughput analyses of interaction between sugarcane and Colletotrichum falcatum, thehemibiotrophic pathogen causing red rot, was made by comparing transcriptomes of compatibleand incompatible interactions in a tropical sugarcane cultivar Co7805 exhibiting differentialreaction to the pathotypes Cf94012 and Cf87012, respectively. Suppression subtractivehybridization (SSH) combined with Illumina 2000 high throughput sequencing was used to identifythe differential transcripts in resistance response library (RRL) and susceptible response library(SRL). After processing and filtering the raw reads from Illumina 2000 high throughput sequencingof subtracted products, we obtained 10,038 and 4,022 high quality transcripts, from RRL and SRL,respectively. Based on the transcripts mapping to KEGG-KASS database, the presence of aCEBiP receptor and the signals ROS, Ca2+, BR, JA and ABA were identified in both theresponses. However, MAPK, ET, PI signals and JA amino conjugation were found only in theincompatible interaction and expression of 10 transcripts involved in these pathways was validatedusing qRT-PCR. Our study concludes that perception of PAMPs occurs in both systems, butdownstream signaling through MAPK, ET, PI and JA amino conjugation and activation of R genesoccurs only in the incompatible interaction. This is the first detailed transcriptomic analysis ofcompatible and incompatible interactions in sugarcane with two different C. falcatum pathotypesthrough SSH and the next generation sequencing (NGS) platform.

Prevalence and RT/RNase H genealogy of Sugarcane bacilliform virus isolates from China

Xiao-Bin Wu, Olufemi J Alabi, Mona B Damaj, Sheng-Ren Sun, T Erik Mirkov, Hua-YingFu, Ru-Kai Chen and San-Ji GaoProceedings of the International Society of Sugar Cane Technologists, volume 29, 2016

Prevalence and genetic diversity of Sugarcane bacilliform virus (SCBV) across major Chinesesugarcane-growing areas were investigated from about 280 sugarcane leaf tissue samplescollected from six provinces in China, 25 from three states in the USA and five from Queensland,Australia. Samples were tested for the presence of SCBV by polymerase chain reaction (PCR)using newly designed degenerate primers targeting a 720-base pair (bp) fragment of the reversetranscriptase/ribonuclease H (RT/RNase H) genomic region. PCR-amplified fragments from 94SCBV-positive samples were then cloned, sequenced and analyzed for their genetic diversity. Theresults revealed considerable haplotype diversity within individual SCBV isolates. Phylogeneticanalysis revealed the segregation of global SCBV isolates into three major monophyletic cladesencompassing 18 subgroups, including five previously undescribed subgroups named as SCBV-Nto -R.

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INTERNATIONAL EVENTS CALENDAR

2018 MEETINGS AND CONFERENCES

April 12: AVH Symposium 2018,Reims, France AVH

May 15-18: ASCPC Conference, Le Pavillon Hotel, New Orleans, LA, USA Contact us

June 5-8: IIRB Congress (International Institute for Beet Research), Deauville, France IIRB.org

June 25-27: ASSCT, Joint Division Meeting, Hyatt Regency Coconut Point, Bonita Springs, FL, USA, ASSCT.org

June 27-29: Sugar Processing Research Institute (SPRI), 2018 Conference,Bonita Springs, FL USA [email protected]

August 3-8: 35th International Sweetener Symposium, The Grand Traverse Resort, Traverse City MI USA, ASA

September 24-28: Association of Latin American Sugar Technologists (ATALAC), Colombia ATALAC

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STORY OF SWEETSi. Motichoor LadooIngredientsBesan, coarse – 1 cup Sugar – 3/4 cupOrange food colour – 1/4 tsp Water – 1 cupMilk – 3 tbsp Cardamom – 1, powderedSalt – a pinch Rose essence – 3 dropsOil/ ghee – to deep fry Lemon juice – 1/2 tspPistachios - 12DirectionsTake besan (kadalai mavu/ chickpea flour) in a mixing bowl, add salt and whisk well or you cansieve too. Add milk, food colour and required water to make a thick batter. Heat a broad kadai with enough oil. Keep a heavy, taller (than the kadai using) box/ dabba nearthe kadai, without touching the flame or the kadai. Once the oil is hot, put flame to low or medium and hold the boondi ladle in slanting way, abovethe oil, so that the handle rests over the dabba. All over the ladle rather than at one place. Once the bubbles reduces and you could see boondis, remove it from oil using a wire mesh ladle.Never let it for more time, otherwise it will get crispy and brown. Drain in paper towel. To make sugar syrup, boil sugar and water until one string consistency. That is, if you swipe theback of the ladle with your fore finger and check between your thumb, a string should form.Switch off the flame, add lemon juice, cardamom and rose essence.Add the prepared boondi, mix well and keep it closed for minimum 20 mins or until all the syrup isabsorbed by the boondi.Chop finely and fry the pistachios in ghee. Add to the soaked boondi, mix.Grease you hand with ghee. Take palm full of the boondi and make ladoos. Repeat to finsish andarrange in serving plate.

ii. Chum ChumIngredients1 liter whole cow milk ½ tsp cardamom seeds50 grams khoya/mava 2 ½ tbsp white vinegar1 ¾ cup sugar for syrup 4 cups water1 ½ tsp fine semolina (sooji) 2 slitted cherries2 tsp sugar Few saffron strands1 tbsp milkDirectionsHeat thick vessel on medium heat and add milk. Now let the milk come to the boil. Once milk willstart boiling turn off the stove and let milk sit for a minute. Line a sieve with muslin cloth/cheese cloth and drain chenna. Pick up cloth and squeeze withlight hand. Make a knot and hang cheese cloth for about 40 minutes.After 40 minutes take out chenna from muslin cloth on clean surface. Chenna will turn smoothand chenna will release its fat and your hand will be greasy. Roll the chenna dough and divideinto nine equal parts. Heat pressure cooker on medium heat and add 4 cups of water and 1 ¾ cup sugar. Stir onceand add cardamom seeds. In boiling sugar syrup gently drop cham cham one by one and closethe lid. Heat should be on medium-high. Wait for 2 whistles and turn off the stove. Now pourthick sugar syrup in which you cooked cham cham and refrigerate it for 30 minutes.Crush saffron strands with 1 tsp of sugar in a mortar. Add 1 tbsp milk and mix well. In a bowltake khoya add saffron, milk and sugar. Mix well and creamy khoya stuffing is ready.Take out cham cham from the refrigerator. Now take out cham cham from the sugar syrup andplace in a plate.

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GUIDELINES FOR AUTHORS

Dear Fellow Author(s),

Pakistan Sugar Journal (PSJ) offers research, analysis, and reviews to keep its local andinternational readership up to date with latest developments in the sugar industry. PSJtakes into account the application of research and focuses on areas in agriculture related tosugar, milling and processing.

In order to have your articles published in the PSJ, you are requested to adhere to thebelow instructions and prerequisites to enable timely review of your submissions by theeditorial board:

I. Write the title of your article in CAPITAL LETTERS in the center of the page.II. Write the complete name of all authors with their addresses – it is compulsory in the

text. References should be cited by author and years as, for one, two or moreauthors (Hammer, 1994, Hammer and Rouf, 1995; Hammer et al., 1993),respectively.

III. Write HEADINGS in bold letters and in the center of the page.IV. Type your article only in ARIAL format.V. Send TABLES and FIGURES on separate page with bold title and mark its numbers

correctly.VI. Observe the following rule for REFERENCE, for one author: Hussain, K. 1991 for

two authors; Khan, M. and A. Habib 1995, for more than two; Ali, K., A.Hussain and S. Nasir, 1990.

VII. Always send two soft copies and one hard copy of CD. Please do not use FLOPPYDISK for this purpose.

VIII. Send copies on an A-4 size page, preferable LASER PRINT in word documentIX. Papers published in the PSJ are free of charges (for authors).X. Send your papers to following address by mail or email:

Dr. Shahid AfghanEditor-in-Chief, Pakistan Sugar JournalShakarganj Sugar Research Institute, Jhang (Pakistan)Phone: +92 47 111-111-765 | Ext. 602, 603Email: [email protected]

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Documents

PAKISTAN SUGAR JOURNAL€¦ · [email protected] ABSTRACT Biomass-based cogeneration in the sugar industry is a useful alternative to meet the shortfall of energy by using