<div style="text-align:justify">Background: Annona spp. are one of the underutilized fruit crop having great commercial and medicinal value and large diversity present in Maharashtra which is untapped. The present study conducted to characterize the Annona genotypes by ISSR and SSR markers for future germplasm conservation and crop improvement programme.&nbsp; 
Methods: Ninety genotypes were collected from various regions of Maharashtra. These ninety genotypes and eleven varieties from All India Coordinated Research Project on Arid Zone Fruits, MPKV, Rahuri were screened for morphological and biochemical characters. After morphological and biochemical characterization twenty-two superior genotypes were characterized using ISSR and SSR markers.&nbsp; 
Result: Among the 29 ISSR primers used, 20 were found polymorphic which produced a total of 171 reproducible amplicons, in which 109 amplicons (75.43%) were polymorphic. In 20 SSR primers used, 14 primers were found polymorphic which produced a total of 43 reproducible amplicons, in which 36 amplicons (83.72%) were polymorphic. Twenty unique amplicons produced by ISSR primer and eight unique amplicons produced by SSR in studied genotypes is useful in genotype/variety identification in future crop improvement. The dendogram generated based on UPGMA method of cluster analysis using ISSR and SSR marker data revealed little different but similar grouping of genotypes into two major clusters viz., cluster A and cluster B. The UPGMA based cluster analysis using dice similarity coefficient grouped Annona genotypes into two major clusters which differentiate squamosa and atemoya species. The distribution of the genotypes in the dendogram was mostly consistent with the known pedigree information, geographical locations and the morphological attributes. Genotypes collected from Purandar, Sawargaon, Pemgiri and Ajanta districts of Maharashtra formed cluster as per geographic collection. The close relationship across genotypes might be explained by either historical relationship to sharing common ancestor or more likely geographical proximity and large population size which favour genetic interchange. This low genetic difference among genotypes suggests that there was more gene flow through random mating without barrier within agro-ecological zones. ISSR and SSR markers gives direction in characterization of Annona genotypes but more areas need to be survey and explore for tapping genetic diversity to widen the gene pool. The screened material found superior need to be prioritized in terms of in-situ and ex-situ conservation for further evaluation and crop improvement.</div>

Characterization of Annona Genotypes by ISSR and SSR Markers

<div style="text-align: justify;">The evaluation and characterization of germplasm of mungbean are considered to be essential prerequisites for a rational use of its genetic resources. Accordingly in the present study, genetic diversity in 18 mungbean genotypes was assessed using a panel of 40 microsatellite based primer pairs. Out of 40 primers initially tested, only 24 showed distinct polymorphism and consequently only these primers were utilized for the purpose of genome profiling of the entries. Altogether, 183 allelic variants were detected with an average of 4.95 polymorphic fragment per primer. The number of amplified loci varied from 4 (CEDG 15) to a maximum of 12 (CEDG 92 and CEDG 172) with an average of 4.95 polymorphic fragment per primer. The polymorphic information content (PIC) ranged from 0.440 to 0.928 with an average of 0.822 per SSR primer pair. The lowest polymorphism percent was observed for primer CEDGAG001 (16.67) while the primer CEDG 154 gave the maximum polymorphism percent (63.64). The primer pairs CEDG 008, CEDG 068 and CEDG 154 among the total primer pairs were found to be highly informative. Dice&rsquo;s similarity coefficient ranged from 0.03 to 0.58. Dendrogram based on SSR data grouped the mungbean genotypes into seven clusters. The pattern of clustering was also reiterated by the results of principal component analysis (PCA). The SSR primer based analysis allowed unique and unambiguous genotyping of the entries. The genetic diversity observed in the present study could be useful to know the phylogenetic links among the cultivars and for the selection of suitable parents to be further used in breeding programmes and genetic mapping studies.</div>

SSR marker based profiling and diversity analysis of mungbean [Vigna radiata (L.) Wilczek] genotypes

<div style="text-align: justify;">Jackfruit (Artocarpus heterophyllus Lam.) being an important dryland horticultural tree, grows well with minimum care and maintenance. The study aimed at identifying the genetic diversity present in twenty jackfruit genotypes for different pulp colours. The genotypes were identified for commercial cultivation, based on fruit and flake quality characters viz., fruit weight, fruit rind weight per kg fruit, number of flakes per kg fruit, weight of flakes per kg fruit, individual fresh flake weight and TSS as per the Jackfruit descriptors given by IPGRI, Rome. The best genotypes identified from the study were Swarna, Lalbagh Madhura, Byrachandra, NSP, Ashoka Yellow and NEL and these can be used for commercial purpose. Molecular diversity analysis was also carried out using 22 SSR primers, out of which six primers (SSR 9, SSR 10, SSR 30, SSR 34, SSR 45 and SSR 48) showed polymorphism among twenty genotypes. The genetic similarity co-efficient ranged from 0 to 0.96, indicating a vast variation in genetic diversity for pulp colour. A dendrogram was constructed by UPGMA analysis revealed three major clusters. The genotypes with cream and yellow colour pulp grouped in cluster I and II, followed by yellow, orange and red in cluster III. These markers effectively segregated the genotypes based on different pulp colours and hence, can be used for both diversity analysis and in breeding applications.</div>

Morphological and molecular genetic diversity analysis using SSR markers in Jackfruit (Artocarpus heterophyllus Lam.) genotypes for pulp colour &nbsp;

<div>Background: Assessing genetic diversity in greengram is a prerequisite for its genetic improvement for yield and quality. DNA markers such as simple sequence repeats (SSRs) have been preferred in this crop for the analysis of genetic diversity because SSR markers are locus-specific, widely dispersed throughout the genome, highly polymorphic due to variation in repeat units, highly informative because of co-dominant nature, high reproducibility and ease of assay by PCR (Polymerase chain reaction). The current study aimed to study the genetic diversity among the mutants of greengram at the molecular level using SSR markers. 
 
Methods: In this study, twenty-five SSR markers were used to analyze the genetic diversity amongst twenty mutant genotypes of greengram along with their parents. Genomic DNA was isolated from the leaves by using the standard CTAB DNA extraction method. Then DNA purification and PCR amplifications were carried out. The genetic variability and diversity among genotypes was examined by assessing the scoring of the amplified bands by SSR -PCR amplification. 
 
Result: Seventeen SSR primers generated 102 polymorphic bands with an average of the six polymorphic bands per primer. The number of alleles per locus ranged from four to nine. The size of amplification product varied for each primer and the range found to be 100bp to 2000bp. The mean polymorphic information content (PIC) value for SSR markers was found to be 0.6335. The value of Jaccard&rsquo;s similarity coefficient had ranged from 0.07-0.70 with an average value of 0.38. The dendrogram constructed on SSR molecular markers data through the unweighted pair group method with arithmetic averages (UPGMA) method had enabled grouping of the genotypes into thirteen clusters. The results indicate the usefulness of SSR markers in the assessment of genetic variability and diversity among the mutant genotypes of greengram.</div>

SSR-marker Assisted Evaluation of Genetic Diversity in Greengram [Vigna radiata (L.) Wilcezk]

Background: Simple Sequence Repeat (SSR) markers are crucial tools for molecular breeding and genetic diversity studies in legumes. Traditional SSR marker development relies on subjective quality assessment methods, which are time-consuming, costly and prone to inconsistency. The lack of quantitative frameworks for predicting marker quality limits the efficiency of breeding programs and genomic studies. To develop a comprehensive machine learning framework for automated SSR marker quality prediction, identify key determinants of marker success through quantitative analysis and establish evidence-based design principles for efficient marker development in legume crops. 
 
Methods: We engineered 15 predictive features from primer design parameters and SSR structural characteristics of 99 horsegram SSR markers. Three machine learning algorithms (Random Forest, Support Vector Regression and Neural Network) were trained and validated using cross-validation. Feature importance analysis quantified the contribution of each parameter to marker quality prediction. 
 
Result: Random Forest achieved optimal performance (R&sup2; = 0.378, MAE = 8.106) with 37.8% explained variance in marker quality prediction. Feature importance analysis revealed primer compatibility factors as dominant predictors: GC content balance between primers (35.6% importance) and melting temperature compatibility (32.2% importance). SSR structural features contributed 22.9% importance, with motif complexity (8.6%) and motif length (8.5%) being most significant. Cross-validation confirmed robust model performance (CV R&sup2; = 0.342 &plusmn; 0.089) across different data subsets.

Machine Learning Framework for Automated SSR Marker Quality Assessment in Horsegram (Macrotyloma uniflorum)

<div>Background: Agricultural research chiefly focuses on the ways to increase productivity of staple food crops like wheat and rice, but still there are crops where research focus is meagre like nutritionally important mungbean crop grown by marginal farmers in crop rotation systems. Mungbean is leguminous crop which is high in protein content thus it offers health benefits at cheaper rates. The present work emphasizes on finding genetic diversity in mungbean germplasm on the basis of chemical and molecular analysis for micronutrients variation (iron and zinc). The identified molecular markers having linkage with high iron and zinc concentrations in the seeds can prove helpful in expansion of biofortification programme. 
 
Methods: Fifty-one green gram genotypes viz. varieties released from CCS Haryana Agricultural University (HAU), Hisar, Punjab Agricultural University (PAU), Ludhiana, Indian Institute of Pulse Research (IIPR), Kanpur and some advanced breeding lines were included in the study. Acid digested samples were used for determination of Fe and Zn by Atomic Absorption Spectrophotometer. Young seedlings leaves were used for isolation of genomic DNA using 2% CTAB (cetyltrimethyl ammonium bromide).&nbsp;&nbsp; &nbsp; 
 
Result: Total of fifty-one mungbean genotypes were tested using fifty simple sequence repeat (SSR) primers. Out of fifty primers screened, 16 primers generated 35 bands. Iron (Fe) and zinc (Zn) in mungbean lines was 36.90 to 107.1 mg/kg and 14.2 to 53.8 mg/kg respectively. The molecular studies based on SSR markers also indicates existence of ample genetic diversity at molecular level.</div>

<div>Genome Analysis of Mungbean [Vigna radiata (L.) Wilczek] using Simple Sequence Repeats (SSR) Mapping Data</div>

<div>Background: Lentil is an important legume crop that plays a vital role in sustainable agriculture and human health. However, the restricted genetic foundation or parentage of contemporary cultivars has arisen as a serious challenge for lentil development. Therefore, determining the genetic diversity and yield-related characteristics is crucial for the breeder to broaden the genetic base and aid in selecting the desirable parents for hybrid development programmes.&nbsp; 
 
Methods: In the current study, microsatellite markers were used for diversity analysis among 37 lentil genotypes. Morphological and molecular systems both were able to differentiate lentil genotypes. Among applied SSR primers 10 were able to produce successful amplifications with template DNA of all the studied genotypes of lentil.&nbsp; 
 
Result: A sum of 357 scoreable bands were produced, 148 of which accounted for 41.45% polymorphism. The UPGMA dendrogram grouped 37 lentil genotypes into 2 groups. PIC values ranged from 0.37 to 0.77. For this experiment, SSR 80, SSR 130, SSR 34-2 and SSR 33 were highly informative polymorphic markers, demonstrating the efficacy and higher resolution in exposing molecular genetic diversity among lentil genotypes. The highest numbers of alleles (5) were produced by SSR 80 primer which was selected as extremely polymorphic. This study reveals the variation across lentil genotypes, which might be employed further in breeding efforts for lentils that result in strong heterosis in the segregating generation. The SSR markers found as polymorphic may also be utilized for further polymorphism analysis among different set of lentil genotypes.&nbsp;</div>

<div style="text-align:justify">&nbsp;</div>

SSR Marker-based Molecular Characterization of Lentil (Lens culinaris Medik.) Genotypes

Background: Cowpea is an important fodder legume crop due to its high protein content and fast growing nature. Cowpea can be grown as sole crop or it can also be grown as intercrop with some non legume such as maize, pearl millet or sorghum. Mostly the farmers prefer erect or semi erect high yielding cowpea varieties as an intercrop because they are easy to harvest. The transfer of erectness along with other forage traits can be done with proper knowledge of gene action controlling that particular trait.&nbsp; 
 
Methods: To improve the fodder yield, two sets of crosses along with their segregating generations were analysed for erectness along with other forage traits to estimate the type of gene action through generation mean analysis. Identification of SSR primers associated with erectness was also carried out in F2 population of a Cross (C-88 x TNFC 6926). 
 
Result: The scaling test showed significant results for most of the traits indicating the presence of epistatic interactions. The six-parametric model test was performed and the results indicated the significant overall mean and traits were inherited quantitatively. The high magnitude of additive x additive gene effect for green fodder yield suggests the pedigree method is most suitable breeding programme for development of fodder cultivars with better quality traits. Out of 151 SSR primers, 15 SSRSs showed polymorphism between two parental lines. Out of these 15 SSR primers, three primers showed linkage with erectness.

Genetic Analysis and Identification of Linked SSR Markers for Erectness in Forage Cowpea [Vigna unguiculata (L.) Walp.]

<div style="text-align: justify;">One hundred cowpea genotypes were screened against Bean common mosaic virus (BCMV) by artificial inoculation genotypes. The genotypes showed different disease reaction viz., immune, resistant, moderately resistant, Sap moderately susceptible, susceptible and highly susceptible. The SSR markers were characterized using a total of 25 randomly selected resistance and susceptible cowpea genotypes. Among, 10 SSR markers used to screen polymorphism, only 4 primers were found to be polymorphic and they were used for characterization and finding out genetic relationship of cowpea genotypes. The polymorphism information content (PIC) of these SSR markers ranged from 0.23 to 0.37 and single marker analysis (SMA) revealed, out of these 4 SSR markers M 135 marker explained 28.2 % of phenotypic variation (R2). The banding pattern of all cowpea genotypes were scored and dendrogram was constructed. A dendrogram of these genotypes based on SSR polymorphism divided into two major clusters at 35 % similarity. Cluster I was the biggest with 20 genotypes which was further separated as three sub-clusters. Whereas, 5 genotypes grouped in cluster II with two small sub-clusters. The clustering pattern of resistant and susceptible genotypes was not clear, however resistant genotypes such as V-5 and IC 8996 clustered in one group with 100 % similarity. The information provided by the identified markers would be very useful in breeding programs to select cowpea lines resistance to BCMV disease and M 135 was found to be good for rapid and accurate screening of cowpea genotypes against BCMV disease resistance.</div>

Characterization and identification of SSR markers for screening of cowpea genotypes against Bean common mosaic virus (BCMV) disease resistance

<div>Lentil is an important pulse crop with high nutritional value and high market price worldwide. Molecular markers have emerged as useful tools to assess the genetic diversity across crops. The study was conducted to explore genetic diversity of twenty lentil genotypes considering yield and yield attributing traits. Among all genotypes, BARI Masur-6, BARI Masur-7 followed by genotypes BD-3806 and BD-4090 showed the highest value of yield attributing traits therefore, these genotypes are considered as best performer. The results of cluster analysis based on the Ward&rsquo;s method grouped the genotypes into three clusters and the genotypes of cluster III revealed the maximum value for yield per plants which indicated their importance in the selection for yield improvement program of lentil. Afterwards, 20 genotypes were evaluated through 7 sets of SSR primers to assess genetic diversity among the genotypes. Among them, four sets of primers viz., SSR 19, SSR 33, SSR 90 and SSR 213 showed high polymorphism which suggesting the greater genetic diversity in the genotypes. The unweighted pair group method of arithmetic mean (UPGMA) dendrogram based on Nei&rsquo;s (1973) genetic distance led the genotypes into four major clusters which showed a bit deviation with the morphological cluster. The findings of this study will be very useful for selection of appropriate parents and the genetic understanding for the set up for future systematic lentil breeding programs.</div>

<div>Morphological and SSR Marker Based Diversity Analysis of Lentil (Lens esculenta) Genotypes using Yield and Yield Contributing Characters</div>