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ThesisItem Open Access Evaluation of wheat genotypes (triticum aestivum) for drought stress tolerance by using morpho-physiological parameters and molecular markers(Vasantrao Naik Marathwada Krishi Vidyapeeth, Parbhani, 2022-02-23) Gormali, Sagar Ashok; Bharose, A.A.Wheat (Triticum aestivum.) is an important cereal crop that provides more carbohydrate and protein than rice. Wheat yields in dry areas have not improved as easily as in more favorable environments or where water is not a limiting factor drought severely reduces wheat productivity in many parts of the world. According to some estimates, approximately 50% of the world's 230 million hectares of wheat are cultivated annually and are regularly affected by drought. The 15 morphophysiological parameter were evaluated of which parameters viz. chlorophyll content, day to heading, spike length, 1000 per grain weight, plant height and grain yield show a large decline percentage under stressed condition than in the control condition. Very less changes under both drought and normal condition was recorded in DBW-150(C). Three molecular marker systems, RAPD, SSR, and ISSR, were used to identify markers associated with drought tolerance in wheat resistance Based on the number of bands and the nature of amplification, 12 RAPD primers, 13 SSR primers, and 10 ISSR primers were selected for the study. The RAPD primer MR-OPX-01 determined to be informative (0.99 PIC value) giving two polymorphic loci with 100% polymorphism and that primer was associated with drought stress tolerance in wheat genotypes. Among 13 SSR primer Xwmc233-5D and Xgwm332-7A showed 100% polymorphism in all 16 wheat genotypes with highest PIC values of 0.77 and 0.75 respectively. The ISSR primers UBC-825 and UBC-841 exhibit the highest levels of polymorphism (83.33%). Each primer generated 3.66 amplicons on average, loci sizes ranged from 150 bp to 1 kb.The SSR primers Xgwm260-7A and ISSR primer UBC-825 exhibits a distinct banding pattern between 700 and 900 bp, and it is possible to confer that this QTL is responsible for the drought stress tolerance in wheat genotypes.Based on assessments of the combined SSR, ISSR, and RAPD marker systems scoring the first, second, and third revealing eigenvectors accounted for (121.12%, 14.87%, and 11.75%, respectively) percent variances with 24.45 percent of the total variation.ThesisItem Open Access Evaluation of linseed (Linum usitatissimum L.) germplasm lines for drought tolerance(Vasantrao Naik Marathwada Krishi Vidyapeeth, Parbhani, 2023-03-03) Jadhav, Akshaykumar Balaji; Dudhare, M.S.Linseed (Linum usitatissimum L.) is a vital oilseed and fibre crop grown for its seed still as fibre which is employed for the manufacture of linen. The oil is edible and even have industrial use like preparation of paints, varnishes, printing ink, soap, oil, cloth, leather, and waterproof fabrics. In the present study the investigation done on linseed morphology and molecular screening for drought tolerance. Under normal conditions among all genotypes earliest flowering was observed in DEEPIKA (41 days). Under stress condition the earliest flowering variety are KARTIKA (41 days) and DEEPIKA (41 Days). Under normal conditions among all lines, the tallest plant was observed in normal condition is RLC-172(55.80cm). Under stress condition the tallest plant was observed is RLC-172 (56.10cm). Under normal conditions the maximum number of fertile capsules observed in RLC-161(24). Under stress condition the maximum number of fertile capsules observed in RLC-143 and RLC- 92(15). Under normal conditions the maximum 1000 seed weight observed in RLC- 167(6.80gm). Under stressed conditions the maximum 1000 seed weight observed in RLC-172(6.15gm). The stressed linseed genotypes showed rise in proline accumulation over unstressed control. Under normal conditions the highest proline content observed in KIRAN (6.62 µmol/g). Under stress condition the proline content the highest proline content observed in KIRAN (18.89 µmol/g). The maximum root length observed in normal condition is KARTIKA(8.6cm). Under stressed condition the root length varies from 3.75cm to 8.96cm with an average of 6.37cm. maximum root length in stressed condition observed in KARTIKA(8.96cm). The present ISSR primers investigation revealed that the mean polymorphic information content (PIC) score for each primer ranged from 0.34 to 0.71 with a mean of 0.55 The least relevant primer was noticed to be ISSR primer UBC-807 of PIC value of 0.34, while the maximum informative marker UBC-825 had a PIC value of 0.71. The similarity matrix of 18 linseed lines varies from 0.20(IA-32-DEEPIKA) to 0.92(RLC-167-RLC-148). Thus, most similar genotypes observed were RLC-167 and RLC-148. The most dissimilar genotypes observed were IA-32 and DEEPIKA were noticed. The dissimilarity matrix of 18 linseed lines varies from 0.08(RLC-167- RLC-148) to 0.77 (IA-32-DEEPIKA). Thus, most dissimilar genotypes observed were IA-32-DEEPIKA. The most dissimilar genotypes observed were RLC-167-RLC-148 were noticed. The present SSR primers investigation revealed that the mean polymorphic information content (PIC) score for each primer ranged from 0.15 to 0.84. The least relevant primer was noticed to be SSR primer LU-9 of PIC value of 0.15, while the maximum informative marker LU-3 had a PIC value of 0.84. Thus, most similar genotypes observed were RLC-92 and RLC-153 and RLC-133. The most dissimilar genotypes observed were IA-32 and KIRAN were noticed. The dissimilarity matrix of 18 linseed lines varies from 0.00(RLC-92-RLC-153) to 0.77 (IA-32-KIRAN). Thus, most dissimilar genotypes observed were IA-32-KIRAN. The most dissimilar genotypes observed were RLC-92 and RLC-153 were noticed.ThesisItem Open Access Molecular characterization of segregating f6 generation of okra (Abelmoschus esculentus (L.) moench)(Vasantrao Naik Marathwada Krishi Vidyapeeth, Parbhani, 2023-02-23) Gawai, Rahul Wasudeo; Bhalerao, S.R.Okra is a nutrient-rich important crop with commercial value in tropical and subtropical regions of the world. In present investigation, eighteen okra genotypes were used and performances of okra genotypes on various yield attributing characters were assessed. Field screening of eighteen okra genotypes for various yield attributing was done. The genotype PBNLF-122 was observed as high yielding genotype that shows better performances viz.,number of branches per plant (1.62), number of nodes per plant(11.65), number of fruit per plant (11.34), fruit weight(12.10) and fruit yield per plant(157.85) whereas genotype PBNLF-116 gave the least performances. Three molecular markers systems, RAPD, ISSR, and SSR, were used to identify genetic relationship among okra genotype. Based on the number of bands and the nature of amplification, 12 RAPD primers, 3 ISSR primers and 6 SSR primers were selected for the study Two RAPD primer OPY-18, OPG-08 showed 100% polymorphism OPY-18 was found to be the most informative marker,withaPICvalueof0.81. UBC 840 were found to be the most informativemarker,withaPICvalueof0.22 and 83.33%. Out of five SSR primersOYVMVwasobservedtobe the most informative with the PIC value 0.77. Genetic similarity was estimated using clubbed data of RAPD, ISSR, and SSR using Jaccard’s coefficient which is ranged from 0.61 (PBNLF-124 and PBNLF-122) to 0.87 (PBNLF-139 and PBNLF-109), with 0.74 being theaverage.The genetic distances ranged from 0.13 to 0.41 with an average 0.27 among these eighteen genotypes of okra. This confirmed that the ultimate genetic distance was maximum in two i.e. 0.41 between PBNLF-122 and PBNLF-104 and second PBNLF-123 and PBNLF-114; The outcome of the field analysisand molecular analysis can be used for hybridization programs and crop improvement program of segregating okra genotypes.ThesisItem Open Access Studies on ems induced mutagenesis in soybean [Glycine max (L.)](Vasantrao Naik Marathwada Krishi Vidyapeeth, Parbhani, 2023-02-15) Kamble, Kranti Pandit; Bhagat, Y.S.Soybean (Glycine max L.) is considered as one of the most nourishing food due to its high protein content which also meets requirements of all the essential amino acid to support normal growth and development of individuals. It also offers economic empowerment to the farmers due to its uses far beyond food and feed. Because of its many uses, it is known as the golden bean and the miracle crop of the 21st century. Mutations play a vital role in any crop improvement programme particularly which naturally occurring genetic variability has exhausted. The genetic variability present in any crop is of vital importance in the formulation of effective breeding programme. Ethyl Methane Sulfonate (EMS) is a mutagenic, teratogenic and possibly carcinogenic organic compound with formula C3H8O3S. It produces random mutations in genetic material by nucleotide substitution; particularly by guanine alkylation and it typically produces only point mutations. Soybean mutant lines treated with different EMS concentration exhibited significant differences for different morphological variations viz., number of primary branches, number of seeds per plant, plant height and number of pods per plant and test weight. Laboratory evaluation of M1 generation treatments viz., M1T2, M1T4 and M1T6 treatments showed significantly higher germination percentage on 21 DAT. M1T6 and M1T9 treatments showed significantly lower number radicle length on 21 DAT, while M1T10, M1T1 and M1T2 showed significantly higher plumule length. In field conditions, the treatments viz., M1T1 and M1T2 showed significantly highest germination percentage on 7 DAS as well as plant stand (%) on 30 DAS. The number of branches was significantly less in number in treatment M1T10. The treatment M1T2 shows significantly higher pods per plant. The seeds per plant and plant height showed significantly less number in M1T9 treatment. In field conditions M2 mutant lines viz., M2T1, M2T7 and M2T8 showed significantly highest germination percentage on 7 DAS, while treatments M2T1, M2T4 and M2T7 showed significantly highest plant stand (%) on 30 DAS. The number of branches was significantly less in number in treatments viz., M2T5 and M2T6. The treatment M2T10 showed significantly less number of seeds per plant. The treatments viz., M2T7 and M2T1 showed significantly higher plant height. The mutagen EMS is capable of producing both types of mutants namely viable and lethal, which is an indication of significant effect on soybean. This clearly indicated the potential of EMS to create considerable variability for further breeding programme. The soybean genotypes were genotyped with 14 SSR markers in order to reveal genetic diversity. The high polymorphic content was reported that ranged from 0.00 to 0.96. The genetic similarity coefficient between ten soybean mutant lines was ranged in between 0.130 to 0.875. Highest genetic similarity (0.857) was found between M2T4 and M2T5, whereas lowest genetic similarity (0.130) was found between M2T9 and M2T3, M2T10 and M2T3, M2T10 and M2T4, M2T10 and M2T6. Cluster analysis revealed soybean mutant lines were separated into three main clusters indicating their diversity. This study showed that the highly significant variation was observed for some of the characters as results of EMS treatments of 0.2%, 0.4% and 0.6%. This clearly indicated the potential of EMS to create considerable variability for further breeding programme. The SSR markers can provide useful tools for mutant characterization, conservation and utilization, as well as for genetic and breeding studies in soybean. Genetic variation among soybean mutants based on SSR analysis could be useful to select parents to be crossed for generating appropriate populations intended for genome mapping and breeding purposes.ThesisItem Open Access Studies on genetic diversity of pigeonpea (Cajanus cajan) fermplasm in response to fusarium wilt tolerance(Vasantrao Naik Marathwada Krishi Vidyapeeth, Parbhani, 2023-02-23) Kshirsagar, Pooja Bharat; Chavhan, R.L.Pigeonpea (Cajanus cajan L.) is a vital legume of the tropical and subtropical regions and belongs to family Leguminoseae. It is commonly known as redgram or tur or arhar. Pigeonpea plays a vital role in food security, balanced diet and subsistence agriculture because of its diverse usages in food, fodder, fuel, soil conservation, and symbiotic nitrogen fixation. India has the largest area (3.9 Mha) under pigeonpea cultivation. India is the origin and primary producer of pigeonpea, accounting for over 63% of total global production and 72% of total area. Fusarium wilt (FW), caused by Fusarium oxysporum f. sp. udum is a common and severe fungal disease occurs in pigeonpea growing areas of India. It causes 30-100% losses in grain yield. The typical wilt symptoms of pigeonpea comprises loss of turgidity, slight inter-veinal chlorosis, internal browning of xylem vessels and a purple band on stem extending upwards from the base. The study was executed to determine the level of genetic variation within pigeonpea germplasm and to document the SSR and ISSR markers linked to Fusarium wilt resistance. Fifteen pigeonpea germplasms were screened with Fusarium oxysporumf. sp. udum pathogen by root dip inoculation method in three replications. The percent wilt incidence was occurred in seedling of pigeonpea germplasm in range between 3-95%. The percent disease incidence was within resistant pigeonpea germplasms was ranged between 0-20%. Whereas, in susceptible germplasms, the disease incidence was ranged between 75-95% and for moderately resistant germplasms, it was observed between 40-50%. Seven pigeonpea germplasms BDN-716, BDN-736, BSMR-736, BSMR-853, ICP-8863, MAL-6, and MAL-13recorded wilt incidence less than 10% and regarded as resistant germplasms. The three germplasms namely, BDN-2, BDN-708, and PAU-881 were validated and categorized as susceptible germplasms with wilt percentage in between 75-95%. The remained germplasms BDN-711, BRG-2, BRG-3, BRG-4, and BRG-5 were also validated and categorized as moderately resistant germplasms showed 40-50% wilt infection to the seedlings. In order to assess genetic variability among 15 pigeonpea germplasms, 10 SSR and 6 ISSR markers were screened across these germplasms. SSR markers were amplified total of 254 bands with an average high polymorphic information content value of 0.75. While six ISSR markers amplified a total of 196 bands with average polymorphic information content value of 0.71. Cluster analysis, based upon genotyping by SSR and ISSR markers using UPGMA, grouped 15 pigeonpea germplasms into 3 major clusters. Cluster I comprised of three susceptible germplasm, while cluster II comprised of ten resistant and moderately resistant germplasms and cluster III comprised of two resistant and moderately resistant germplasms. The data generated during disease screening test and genetic diversity studies by using SSR and ISSR marker revealed correlation with each other and showed similar type of clustering pattern generated between two independent studies. This endorse the future utility value of SSR, ISSR markers, used for molecular characterization. The SSR marker UBC-850 showed tight association with Fusarium wilt resistant gene whereas, ISSR marker ISD-32 exhibited close association with Fusarium wilt susceptible gene and significantly differentiated all pigeonpea germplasms according to the Fusarium wilt disease reactions. The present study helps in finding out feasibility of screened SSR and ISSR markers in genetic diversity analysis and their potential association with disease resistance and future applicability in Fusarium wilt resistance breeding program.ThesisItem Open Access Studies on its-rdna based molecular characterization of pseudocercospora species complex in banana (Musa sp.)(Vasantrao Naik Marathwada Krishi Vidyapeeth, Parbhani, 2023-02-15) Kachare, Vaibhavi Anand; Chavhan, R.L.Banana (Musa spp.) is a significant horticultural crop growing in the world. It is the most delicious fruit which is eaten both in cooking as well as table purpose. Banana plants are more susceptible to different diseases due to their limited genetic diversity. The Sigatoka disease, caused by Pseudocercospora species, is damaging the banana plantations in Maharashtra and other states of India that grow banana. Studieswereconductedwiththeaim to identify predominant species among various member species of Pseudocercosporaacross various banana growing locations in Maharashtra and nearby state. Total 16 isolates of Pseudocercosporaspecies were, collected, purified, identified and maintained. The morphological of the species was confirmed with the support of Agharkar Research Institute, (ARI), Pune. Among 16 isolates of Pseudocercospora species, the member species P. musicola was found dominant in banana growing regions of Maharashtra. The in-vitro pathogenicity test of member isolates of Pseudocercospora spore species was executed into the Laboratory. Further to characterize Pseudoceercospora species at nuclear r-DNA sequence level, three representative isolates of the species were sequenced, The nuclear rDNA sequence of the species P. musicola isolate (RLC-1) and P. fijiensis isolate (RLC-4, RLC-8) were annotated and confirmed through nucleotide BLASTn analysis. The rDNA sequence of species P. musicolaisolate (RLC-1) and P. fijiensisisolate (RLC-4, RLC-8) were found 507, 547, 562 bp respectively. At present, information on phylogenetic relationship among Pseudocercosporaspecies of sigatoka disease in banana is lacking. Therefore, anattempt has been made and assessed nuclear rDNA based phylogenetic relationship among Pseudocercosporaspecies of sigatoka disease of banana The nuclear rDNA was employed through insilico as well as real time restriction digestion by hexa cutters and rDNA sequence for phylogenetic analysis studies The dendrogram analysis based upon ribotyping data revealed two distinct clusters. The genetic variability among the species was ranged between 0 to 88.19%. While, genetic similarity was found in the range between 12 to 100 %. Restriction Enzyme BtsIMutI can be utilized as Pseudocercospora species-specific RFLP marker which could specifically discriminated species Pseudocercospora from rest of banana pathogens. The phylogenetic relationship was established by using Neighbour joining method and Clustal omega tool revealed the close association of members of species P. musicolaand P. fijiensis. It also revealed that, two species of Pseudocercospora was found associated with sigatoka disease of banana in Maharashtra and Telangana would have P. musicolaas common evolutionary root for sigatoka disease.ThesisItem Open Access Production of nutrabeverage by fermentation of mahua flower (madhuca indica) Extract(Vasantrao Naik Marathwada Krishi Vidyapeeth, Parbhani, 2022-02-23) Tompe, Takshashila Harish; Surve, V.D.The term ‘nutrabeverage’ was applied to the product with increased amount of nutrients that was produced by alcoholic fermentation using yeast strains, in which sugar was converted into alcohol and carbon dioxide. Mahua flower basednutrabeverageprovides all the therapeutic nutritional benefits of mahua flowers and further, enriches during the process of fermentation.The release of amino acids and other nutrients from yeast during fermentation increases the nutritional value of nutrabeverage. The mahua flowers are nutritionally balanced as a rich source of proteins, carbohydrates, dietary fibers in addition to minerals like calciumand iron. In this study nutrabeverage was prepared from dried mahua flower extract, the juice was filtered and transferred to the fermenter and Saccharomyces cerevisiae was used an inoculum and allowed for fermentation. The effect of physicochemical characteristics of flower extract on fermentation process as well as process conditions were optimized through sensory and nutritional analysis of the nutrabeverage. It was concluded that the optimal fermentation process conditions such as temperature, pH, incubation period and inoculum concentration for the production of nutrabeverage were 30°C, 4.5, 72 hrs and 6% respectively. Nutrabeveragewas tested through sensory evaluation against commercial control as well as nutrabeverage produced at different fermentation conditions. The nutrabeverage produced from raw mahua flower extract at temperature 30°C, pH 4.5, incubation period of 72hrs and 6% inoculum concentration was accepted with respect to sensory qualities like color, taste, flavor, clarity and overall acceptability. It was concluded that mahuaflower as a substrate has potential for making good quality fermented product.ThesisItem Open Access Characterization of linseed (Linum usitatissimum L.) germplasms(Vasantrao Naik Marathwada Krishi Vidyapeeth, Parbhani, 2023-02-15) Kamble, Abhishek Suryakant; Dudhare, M.S.The experiment on titled "Characterization of Linseed (Linumusitatissimum L.) Germplasms" was carried out with the main goal of studying the variability present among the linseed accessions at morphological, biochemical and molecular levels. Eighteen (18) linseed germplasms were evaluated in rabi 2021-2022 at the Department of Plant Biotechnology, VilasraoDeshmukh College of Agricultural Biotechnology, Latur. Analyses for genetic variability, heritability, genetic advance, correlation coefficient, path coefficient, and genetic diversity were performed using morphological, biochemical and molecular characterization. Observations were made on five randomly selected competitive plants from each germplasm in each replication. Analysis of eight character under study viz., days to first flower, days to 50% flowering, colour of flower, plant height at maturity, number of primary branches per plant, number of capsules per plant, number of seeds per capsule, 1000 seed weight. First flower observed in KARTIKA (36 day), late first flower observed in RLC-165 (46). For the character days to 50% flowering, the range was observed from 48 days (RLC-153) to 60 days (RLC-167) with a mean of 53.43 days. Maximum number of capsules per plant observed in DEEPIKA (37), minimum number of capsules per plant observed in R-552 (10.67), tallest plant height observed in RLC-172 (55.17cm) and lowest plant height recorded in R-17 (33cm). Maximum number of seeds of the capsule was found in RLC-148 (10 seed/capsule) genotype while the minimum no of seeds in the capsule was observed in IA-32 (7 seed/capsule). KIRAN (4.32gm) linesrecorded lowest test weight and RLC-172 (6.93 gm) lines show the highest test weight. Maximum number of primary branches recorded in DEEPIKA (7) and lowest in R-552 (2). RLC-133 show white in colour and remaining 17 lines shows in blue colour. Total seed protein profiling of the germplasms SDS-PAGE shows high level similarity between them. According to the similarity index, the dissimilar germplasm (R-7-RLC-161) has a 25% similarity. The current SSR primers investigation revealed that germplasms IA-32 and KIRAN (22%) were the most diverse, and primers LU-3 (0.84) and LU-8 (0.77) demonstrated the highest PIC value and 100% polymorphism. ISSR analysis revealed that genotypes IA-32 and DEEPIKA (20%) were the most diverse, with primers UBC-825(0.71) and UBC-850 (0.70) showing the highest PIC value and 100% polymorphism.ThesisItem Open Access Studies on coat protein gene based molecular characterization of tomato mosaic virus (tomv) infecting tomato (Solanum lycopersicum L.)(Vasantrao Naik Marathwada Krishi Vidyapeeth, Parbhani, 2023-02-15) Lokewar, Shivani Devidas; Hinge, V.R.Tomato (Solanum Lycopersicum L.) is one of the most important vegetable crops belonging to the family Solanaceae and ranks second in terms of consumption after potatoes.Among the viral diseases infecting Tomato, Tomato mosaic disease is one of the most destructive diseases and causes very high yield loss. The present study is conducted on coat protein gene-based molecular characterization of the tomato mosaic virus.The 48 symptomatic samples were collected from different regions of Maharashtra. The coat protein gene-specific primers were used to detect the presence of infection of the Tomato mosaic virus among the collected samples. PCR Thermocycler successfully amplified the 479bp amplicon of the coat protein gene from 17 samples amongthe collected 48 samples. The coat protein gene (479bp) of representative ToMV isolates i. e. ToMV Pune-1, ToMV Parbhani-1, ToMV Latur-2, and ToMV Solapur-1 was sequenced. The BLASTn analysis of four isolated ToMV coat protein gene sequences were showing the highest similarity i.e.,99.74 % with other strains of ToMV at the nucleotide level. In BLASTp analysis, protein sequences showed98.74% similaritywith other coat protein gene sequences of the tomato mosaic virus available in the NCBI database.All four isolated sequences were showing 82.28 to 88.68 % similarity with otherrelated tobamovirusesi. e. Tomato mottle mosaic virus, Tomato brownrugosefruit virus, Tobacco mosaic virus and Chilipeppermildmottlevirus. Among four isolates of ToMV coat protein gene sequences the ToMV Latur-2 and ToMV Solapur-1 sequences showed the highest similarity of 98.95 % at the nucleotide level. At the protein sequence level, it was showing 98.74 % similarity. The ToMV Pune -1 isolate was found diverse from the other three isolates i. e. ToMV Parbhani-1, ToMV Solapur-1 and ToMV Latur-2 at nucleotide sequence (97.04 %) and protein sequence (94.96 %) levels. The highest genetic distance was observed between the ToMV Pune-1 isolate and the other three isolates. ThisindicatedthattheToMV Pune-1 isolates differedfrom the otherthreeisolates collected from Latur, Solapur and Parbhani regions. This confirmed the effect of geographical location and climatic conditions on the structure of coat protein gene evolution. Thecoat protein genes of four sequenced ToMV isolates and related 10 coat protein gene sequences; 4 sequences of ToMV from databases & one each of 6 tobamoviruses (Tomato mottle mosaic virus, Tobacco mosaic virus, Tomato brown rugose fruit virus, Tobacco mild green mosaic virus, Tobacco latent virus and Cucumber green mottle mosaic virus) were utilized for studying phylogenetic analysis. The selected isolates under study were categorized into III clusters at the nucleotide level and II clusters at the protein level. The Tomato mosaic virus is maintaining its separate cluster atthe nucleotide as well as protein sequence level. Among all the related sequences selected for the study Tomato mottle mosaic virus(ToMMV) is found closely related to ToMV and grouped in the same cluster. Thus, in the genus tobamovirus ToMV and ToMMV species were most closely related to each other and maintained their separate species identity. The tobacco latent virus was out-grouped and recorded the highest genetic distance i.e. 2.363 at the nucleotide sequence level and 7.431 at the protein sequence level from ToMV in the tobamovirus genus. The Percent identity within the coat protein gene of ToMV isolates varied from 89 to 100 % at the nucleotide level and at 82 to 100 % at the protein level. Whereas percent identity among the ToMV and other related tobamoviruses ranges from 46 to78% at nucleotide and 12 to 74% at the protein sequence level. The percent identity results confirmed the high variation of coat protein gene sequence at Tomato mosaic virus species level and within the tobamovirus genus. The coat protein gene sequence has high variation at the protein sequence level as compared to the nucleotide sequence level. The variation in the sequence of the coat protein gene is a part of mutation, evolutionary changes and virus evolution. The coat protein gene is playing important role in ToMV virus infection and involved in the cell-to-cell movements, long-distance movements and replication of viruses. Thus variation, mutation and evolution in the sequence of the coat protein gene is an important mechanism of virus adaptation and may lead to the development of new species.
