Sorghum (Sorghum bicolor L. Moench) is fifth mosteconomically important cereal crop of the world. Sorghum is a multipurposecrop, its uses extend from being an important source of food, feed, and forage,it supplies raw materials for industrial use. Sorghum has diverse germplasmgrouped into sweet, forage, energy and grain sorghum types depending on itsuses.
Knowledge of genetic diversity helps breed important crop varieties.Sorghum germplasm has been explored via morphological, biochemical, proximateand molecular markers. Biotechnological methods like axenic culture,transformation, molecular breeding, genomics and proteomics have beensuccessfully utilized in sorghum improvement. Documenting the crop germplasmwith morphological traits variability is considered less efficient asenvironmental conditions influence the results.
Whereas, DNA markers provide anaccurate alternative for assessing the similarities and differences amonggenotypes of any crop germplasm. This study deals with estimation of diversityof high biomass sorghum germplasm procured from USDA. The morphological datawill be used to screen the proficient genotypes, which will be later analysedby microsatellite markers. The study will help identify sorghums havingpotential for breeding of improved energy sorghum varieties in Pakistan.
UNIVERSITYOF AGRICULTURE, FAISALABAD Centre ofAgri. Biochemistry and Biotechnology(Synopsisfor M.Phil. Biotechnology) I.
Title: Understanding genetic diversity of sorghumusing quantitative traitsII. (a) Date of Admission : 22-09-2016(b) Date of Initiation : 22-09-2016(c) Probable Duration : 4 semesters III. Personnel:(a)Name of the Student : MuhammadImran Farooq(b) Registration Number : 2016-ag-2144(c) Supervisory Committee: i) Dr.
Bushra Sadia (Supervisor)ii) Dr. Faisal Saeed Awan (Member)iii) Dr. Jafar Jaskani (Member) IV.
Need forthe Project(Sorghumbicolor L. Moench) is the world’s fifth most vital product after wheat, rice,maize and grain (Motlhaodi et al., 2014). It has a place with the familyPoaceae and the variety Sorghum with a few animal categories and sub-species.It is notable C-4 trim and developed in warm and parched climatic regions ofthe world. It is an imperative sustenance, nourish and scavenge edit. In Africaand Asia, 80% of the overall sorghum is developed as an essential sustenanceand the lingering 16-20% is being developed in the propelled conditions of theworld as feedstuff.
There is an extensive variety of hereditary varietyintroduce in sorghum (Hariprasanna and Patil, 2015). In addition being a hugegrain edit, it gives crude material to the amalgamation of starch, fiber,dextrose syrup, biofuels, liquor, and different items. (Li et al., 2010).
Sorghumis being developed as a grain edit in Pakistan. It is developed on a territoryof 171,000 hectares with generation of 103,000 tons and normal yield of 26.9tons for each hectare (Sher et al., 2016). Conventionalbreeding systems have brought about the successful improvement of highyielding, profoundly altered sorghum cultivars. Be that as it may, it has a fewimpediments because of normal sexual contrariness boundaries and the tighthereditary variety in sorghum.
Consequently, facilitate advancement ofgeneration, quality and protection from biotic and abiotic focuses on needsearnest consideration. Morphological, biochemical and sub-atomic procedureshave been misused for surveying these assets. Biochemical markers then againuncover all the more really the hereditary fluctuation, as they are theimmediate results of qualities. (Iqbal et al., 2010).
Theimprovement in subjective and quantitative attributes of sorghum expandsbiomass highlights and biofuel generation which significantly diminishes thefuel inadequacy (Iqbal et al., 2010). Hereditarydeviation can be surveyed by distinguishing the gatherings which havecomparative genotypes for the evaluation and safeguarding. Evaluation ofhereditary assorted variety is noteworthy to cultivar change and for decidingparticular phenotypes and genotypes.
In any reproducing program hereditaryassorted variety assumes critical part. Hereditary examination and DNA fingerprintingcan be viably considered by DNA based atomic systems (Geleta et al., 2006). Hereditaryassorted variety can likewise be controlled by morphological markers.
Thesemorphological markers are regularly affected by ecological variables,accordingly the data got isn’t extremely dependable (Shehzad et al., 2009). DNAmarkers are getting to be plainly solid device to examine decent variety andcontrol agronomic characters for the change of sorghum cultivars. These markersare useful in enhancing reproducing programs through various ways. Hereditarystocks and cultivars can be recognized by hereditary fingerprints. Reproducersare presently permitted to assess the decent variety and hereditary relatednessby looking at uncommon hereditary signs in germplasm.
Quantitative quality loci(QTL) for some, mind boggling characters can be assessed by utilizingsub-atomic markers (Ejeta and Knoll, 2007). The little and altogether genome ofsorghum makes it a model reap for the use of genomics-based reproducing systems(Batley and Edwards, 2007). Distinctivesorts of atomic markers have been effectively utilized for the appraisal ofhereditary assorted variety in sorghum genotypes. Among these, Simple SequenceRepeats (SSRs) are to a great degree valuable inalienable markers attributableto their primary legacy, high polymorphism and reproducibility. In like manner,ESTs are potentially silly reason for SSRs that uncover polymorphisms not justin the source taxon, but rather in related taxa, also. Objectives: • Geneticdiversity analysis of sorghum genotypes using morphological markers • Geneticdiversity analysis of selected sorghum genotypes using SSR markers • Selectionof promising sorghum genotypes V. Review of Literature Brown et al.
(2011) applied structure and principal componentsanalysis for genetic evaluation based onphysical appearance. In order to check the genetic similarities onthe basis of physical appearance, 434 SNP and SSR allele were genotyped from216 sorghum lines. The structure and principal component study aided in bettercharacterization.Burow et al. (2012) performed a study to investigate sorghum landraces that were 159 collected from regionsof china that had a cool temperature by means of 41 SSR markers. These resultsindicated that 40 out of 41 SSRs were polymorphic and extremely useful.
The PICwas in a normal range of 0.04-0.91.
The genomic resemblance coefficientspredictable range was 0.4-0.9 that indicated variation in sorghum lines.Olwenyet al. (2014) conducted an experiment to check the hereditary uniquenessand correlation among various sorghum genotypes from various nations byutilizing straightforward arrangement rehashes markers. 11 microsatellites wereutilized to identify 86 sorghum types.
Results indicated 8 alleles per locuswith 86 alleles in total. The scored PIC value was 0.53, that coordinated thearrangement of direct decent variety 0.
09~0.89.Adugna (2014) studied 8 land races of sorghum for estimation of thehereditary pattern and in-situ assortment of around 8 sorghumlandraces. They exploited 12 exceptionally polymorphic SSRs of sorghum and 7phenotypic traits for this work. The study indicated high variation amongphenological traits of different sorghums. They reported 123 alleles, out ofwhich 78 were novel. Each loci accommodated on average 10.25 alleles, generatedby twelve microsatellites.
The normal heterozygosity and hereditary estimation rangedfrom 0.04-0.33. Mofokeng et al.
(2014) utilized SSR markers to assesshereditary deviation in sorghum genotypes. 103 landraces and reproducing lineswere genotyped by utilizing 30 SSR preliminaries. The allele size ran from 90to 294bp indicated hereditary deviation in the sorghum. The quantity of alleleswas in the scope of 2-15 with mean of 6.4 for each locus. The mean polymorphicdata content was 0.5031 with the mean heterozygosity of 0.
5483. Aminon et al.(2015) contemplated the hereditary uniqueness in sorghum utilizing 142increases gathered from North Benin and described utilizing 10 subjective and14 quantitative characteristics. More extensive inconstancy were appeared amonggrain yield (0.72-10.57 tons/ha), panicle weight (15-215.
95 g), days to halfblooming (50-195 days), and plant tallness (153.27-636.5 cm). The outcome willexpand the creation and decent variety of sorghum.
Ryu et al.(2016) gathered high-seed-yield sorghum from 3 nations to assess the assortmentand concoction examination. By utilizing 10 SSRs to assess hereditary decentvariety distinguished 37 alleles having 2-7 alleles for every locus.
Threeprimary arrangements of sorghum cultivars were made based on phylogeneticexamination by SSR markers. These gatherings were then contrasted and thosegatherings in light of substance organization. Real fluctuations were found inthe whole grain and substance of all cultivars. Muui et al.(2016) utilized 44 landraces of sorghum from Kenya to assess the hereditaryvariety of sorghum. By utilizing 20 SSR markers 4 assortments out of 44 werebroke down.
The normal estimation of hereditary assortment accomplished was 0.35.The allelic array was 4-12 with normal estimation of 6.
05. Investigation ofatomic deviation demonstrated more noteworthy deviation inside populaces thanamong the gatherings. Sinha and Kumar(2016) inspected forty distinctive sorghum promotions for hereditary decentvariety utilizing quantitative attributes. Perceptions were recorded on 14quantitative attributes, out of which 9 distinct qualities demonstrated highvariability were chosen for hereditary assorted variety examination. Silva et al.(2017) utilized 100 sorghum increases gathered from the germplasm bank of theEmbrapa Maize and Sorghum rearing project to ponder the phenotypic and atomicassorted variety of sorghum. Morphological characteristics identified withsugar and biomass generation were utilized for phenotypic characterizat.
Lowconnection (0.35) was seen amongst phenotypic and sub-atomic decent varietygrids. The outcome showed complementarity between the sub-atomic and thephenotypic characterization to help a breeding system. VI. Material and Methods The planned research study willbe carried out in Somatic Cell Genetics Laboratory, Centre of AgriculturalBiochemistry and Biotechnology (CABB), University of Agriculture, Faisalabad.Plant Material and Cultivation:The sorghum germplasm used in thestudy are provided by Agriculture Research Service, United Sates Department ofAgriculture (ARS-USDA), USA. These varieties will be screened based on theirbiomass related traits. Then diversity analysis of these varieties will be doneby using molecular markers.
Lay out and Phenotyping:The Sorghum germplasm trial willbe sown at the research field area of University of Agriculture, Faisalabad, byusing Randomized Complete Block Design (RCBD) with three replications usingdibbler method. The row × row and plant × plant distances will be 30 cm and 10cm respectively. Fifteen plants will be planted in a line. Identified best performingplants from each genotype per replication will be selected on biomass relatedtraits. Biomass related traits: 1.
Germination Percentage (%)2. No. of Leaves per Plant (n)3. No. of Nodes (n)4. Days to 50% Flowering5. Brix Value6. Fresh Biomass (g)7.
Dry Biomass (g)8. Days to Maturity (n) 9. Plant Height (cm) Molecular Analysis: 1. The fresh leaf samples will beused for DNA extraction following the method reported by Khan et al.(2004).2.
The DNA quantification will be optimized 3. The genetic divergence of sorghum germplasm will beassessed by SSR. Statistical Analysis: The data will be analyzed using POPGENE 32 computersoftware (ver. 1.44) (Yeh et al., 1999) and the efficiency of SSR markers regarding no. of loci, no.
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