Trifolium alexandrinum

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D R Malaviya - One of the best experts on this subject based on the ideXlab platform.

  • phenotype study of multifoliolate leaf formation in Trifolium alexandrinum l
    PeerJ, 2021
    Co-Authors: D R Malaviya, P Kaushal, Ajoy Kumar Roy, Shalini Pathak, Ruslan Kalendar
    Abstract:

    Background The genus Trifolium is characterized by typical trifoliolate leaves. Alterations in leaf formats from trifoliolate to multifoliolate, i.e., individual plants bearing trifoliolate, quadrifoliolate, pentafoliolate or more leaflets, were previously reported among many species of the genus. The study is an attempt to develop pure pentafoliolate plants of T. alexandrinum and to understand its genetic control. Methods The experimental material consisted of two populations of T. alexandrinum with multifoliolate leaf expression, i.e.,interspecific hybrid progenies of T. alexandrinum with T. apertum, and T. alexandrinum genotype Penta-1. Penetrance of the multifoliolate trait was observed among multifoliolate and trifoliolate plant progenies. In vitro culture and regeneration of plantlets from the axillary buds from different plant sources was also attempted. Results The inheritance among a large number of plant progenies together with in vitro micro-propagation results did not establish a definite pattern. The multifoliolate leaf formation was of chimeric nature, i.e., more than one leaf format appearing on individual branches. Reversal to normal trifoliolate from multifoliolate was also quite common. Penetrance and expression of multifoliolate leaf formation was higher among the plants raised from multifoliolate plants. Multifoliolate and pure pentafoliolate plants were observed in the progenies of pure trifoliolate plants and vice-versa. There was an apparent increase in the pentafoliolate leaf formation frequency over the years due to targeted selection. A few progenies of the complete pentafoliolate plants in the first year were true breeding in the second year. Frequency of plantlets with multifoliolate leaf formation was also higher in in vitro axillary bud multiplication when the explant bud was excised from the multifoliolate leaf node. Conclusion Number of leaflets being a discrete variable, occurrence of multifoliolate leaves on individual branches, reversal of leaf formats on branches and developing true breeding pentafoliolates were the factors leading to a hypothesis beyond normal Mendelian inheritance. Transposable elements (TEs) involved in leaf development in combination with epigenetics were probably responsible for alterations in the expression of leaflet number. Putative TE's movement owing to chromosomal rearrangements possibly resulted in homozygous pentafoliolate trait with evolutionary significance. The hypothesis provides a new insight into understanding the genetic control of this trait in T. alexandrinum and may also be useful in other Trifolium species where such observations are reported.

  • complementary gene interaction and xenia effect controls the seed coat colour in interspecific cross between Trifolium alexandrinum and t apertum
    Genetica, 2019
    Co-Authors: D R Malaviya, P Kaushal, A Yadav, D K Pandey
    Abstract:

    Trifolium alexandrinum (Egyptian clover) is a widely cultivated winter annual fodder. Present work deals with inheritance of the seed coat colour in segregating progenies of the interspecific cross between T. alexandrinum and T. apertum. Although, both the parent species possessed yellow seed coat, the F1 seeds were black coloured in the reciprocal cross (T. apertum × T. alexandrinum). Seeds borne on individual F2 plants and the advancing generations segregated in yellow and black seed coat colour, which confirmed xenia effect. F2 seeds collected from individual F1 plants exhibited nine black and seven yellow segregation ratio. The segregation of the seed coat colour recorded from F3 to F5 generations revealed that yellow seed coat was true breeding (i.e. non-segregating) in this interspecific cross (including the reciprocal crosses). However, the black seeded progenies were either true breeding or segregated in nine black: seven yellow ratio or three black: one yellow ratio suggesting a complementary gene interaction or duplicate recessive epistasis. It indicated that the seed coat colour is controlled by complementary gene interaction along with xenia effect in interspecific crosses between T. alexandrinum and T. apertum. Occurrence of the complementary genes across the species could suggest T. apertum to be the progenitor of T. alexandrinum. Inheritance of seed coat colour in reference to its importance in Egyptian clover breeding is also discussed.

  • development characterization and cross species transferability of genomic ssr markers in berseem Trifolium alexandrinum l an important multi cut annual forage legume
    Molecular Breeding, 2015
    Co-Authors: Priyanka Verma, D R Malaviya, P Kaushal, Ajoy Kumar Roy, Amaresh Chandra, Dinesh Pandey, Sabhyata Bhatia
    Abstract:

    Egyptian clover (Trifolium alexandrinum), a multi-cut forage legume, is widely grown as an annual feed crop. Genetic improvement in this legume is largely hampered due to limited availability of genomic resources. Hence, a genomic library enriched for GA/CT motif was constructed to develop SSR primers. As a result, 667 SSR primer pairs were developed and 111 were validated in T. alexandrinum cv. Wardan s-99-1. Among these, 45 primer pairs were used for the analysis of genetic diversity among 86 Trifolium accessions, resulting in 220 alleles with an average of 4.88 alleles per locus. Polymorphic information content for these markers ranged from 0.040 to 0.819 with an average of 0.445. The observed and expected heterozygosity for individual loci ranged from 0.39 to 0.89 and 0.368 to 0.80 with an average of 0.675 and 0.659, respectively. The markers exhibited varied levels of cross-species transferability ranging from 93.33 % across T. resupinatum to 40.0 % across T. balansae. Phylogenetic relationships were analyzed using the unweighted pair group method with arithmetic averages (UPGMA)-based dendrogram which was able to establish the uniqueness of each genotype and grouped them into two major clusters. To the best of our knowledge, the SSR markers reported here are among the first to be reported in this important yet neglected forage legume. They were assessed to be highly efficient and polymorphic and found to be useful for distinguishing the closely related genotypes of Trifolium. This large resource of SSR markers would therefore facilitate new avenues for T. alexandrinum genomics.

  • isozyme variability in Trifolium alexandrinum accessions
    Indian Journal of Genetics and Plant Breeding, 2008
    Co-Authors: B Kumar, D R Malaviya, A K Roy, P Kaushal
    Abstract:

    The present investigation was carried out on 65 accessions of berseem (Trifolium alexandrinum) to estimate the genetic variability based on five enzymes – peroxidase, acid phosphatase, esterase, super oxide dismutase and glutamate oxalo acetate transaminase using starch gel electrophoresis. A similarity matrix showed 95–100% similarity among 64 accessions. Accession EC 329299 belonging to ‘Saidi’ ecotype showed 79% similarity with other accessions. The accessions grouped in three major clusters based on the cutting points in the dendrogram at root level. However, phenetic line drawn at 98% identified 25 groups comprising of 1 to 10 accessions. Of these, 13 groups with more than one germplasm lines showed 100% similarity. Morphologically distinct accessions (i.e. multifoliates and red flowered) formed one cluster. Monomorphic bands (13 out of 26) were in high number. Exotic accessions also showed close similarity to many indigenous accessions which indicated less genetic variability among the germplasm lines introduced from other countries. Polymorphism was recorded for 13 bands (2 for SOD, 3 for GOT, 1 for ACP and 7 for Peroxidase).

  • Trifolium alexandrinum t resupinatum interspecific hybrids developed through embryo rescue
    Plant Cell Tissue and Organ Culture, 2005
    Co-Authors: P Kaushal, B Kumar, D R Malaviya, Ajoy Kumar Roy, A Tiwari
    Abstract:

    This is the first report on the development of interspecific hybrids between T. alexandrinum and T. resupinatum, using embryo rescue. T. resupinatum is an important donor species for traits such as resistance to root and stem rot diseases, and tolerance to alkalinity for the potential improvement of T. alexandrinum. T. alexandrinum, as female parent, was crossed with T. resupinatum and embryos were excised 10–12 days after pollination and cultured in vitro on EC3 medium. Of the recovered embryos, 23.08% showed plumule emergence. For subculturing, shoot-induction medium LSP3 was used. Roots were induced on RL1 medium. Regenerants were hardened and inoculated with Rhizobium before transfer to the field. Morphological appearance of the hybrids was intermediate to both the parents. The hybrids were erect, leafy with short internodes and possessed distinct dwarf bushy compact appearance and were profusely branched. The plants were late flowering with pollen fertility more than 90%. Three out of five hybrid plants set seed following selfing. Hybridity of the plants was also confirmed through Est, PRX, SOD, GOT isozymes and protein banding patterns.

P Kaushal - One of the best experts on this subject based on the ideXlab platform.

  • phenotype study of multifoliolate leaf formation in Trifolium alexandrinum l
    PeerJ, 2021
    Co-Authors: D R Malaviya, P Kaushal, Ajoy Kumar Roy, Shalini Pathak, Ruslan Kalendar
    Abstract:

    Background The genus Trifolium is characterized by typical trifoliolate leaves. Alterations in leaf formats from trifoliolate to multifoliolate, i.e., individual plants bearing trifoliolate, quadrifoliolate, pentafoliolate or more leaflets, were previously reported among many species of the genus. The study is an attempt to develop pure pentafoliolate plants of T. alexandrinum and to understand its genetic control. Methods The experimental material consisted of two populations of T. alexandrinum with multifoliolate leaf expression, i.e.,interspecific hybrid progenies of T. alexandrinum with T. apertum, and T. alexandrinum genotype Penta-1. Penetrance of the multifoliolate trait was observed among multifoliolate and trifoliolate plant progenies. In vitro culture and regeneration of plantlets from the axillary buds from different plant sources was also attempted. Results The inheritance among a large number of plant progenies together with in vitro micro-propagation results did not establish a definite pattern. The multifoliolate leaf formation was of chimeric nature, i.e., more than one leaf format appearing on individual branches. Reversal to normal trifoliolate from multifoliolate was also quite common. Penetrance and expression of multifoliolate leaf formation was higher among the plants raised from multifoliolate plants. Multifoliolate and pure pentafoliolate plants were observed in the progenies of pure trifoliolate plants and vice-versa. There was an apparent increase in the pentafoliolate leaf formation frequency over the years due to targeted selection. A few progenies of the complete pentafoliolate plants in the first year were true breeding in the second year. Frequency of plantlets with multifoliolate leaf formation was also higher in in vitro axillary bud multiplication when the explant bud was excised from the multifoliolate leaf node. Conclusion Number of leaflets being a discrete variable, occurrence of multifoliolate leaves on individual branches, reversal of leaf formats on branches and developing true breeding pentafoliolates were the factors leading to a hypothesis beyond normal Mendelian inheritance. Transposable elements (TEs) involved in leaf development in combination with epigenetics were probably responsible for alterations in the expression of leaflet number. Putative TE's movement owing to chromosomal rearrangements possibly resulted in homozygous pentafoliolate trait with evolutionary significance. The hypothesis provides a new insight into understanding the genetic control of this trait in T. alexandrinum and may also be useful in other Trifolium species where such observations are reported.

  • complementary gene interaction and xenia effect controls the seed coat colour in interspecific cross between Trifolium alexandrinum and t apertum
    Genetica, 2019
    Co-Authors: D R Malaviya, P Kaushal, A Yadav, D K Pandey
    Abstract:

    Trifolium alexandrinum (Egyptian clover) is a widely cultivated winter annual fodder. Present work deals with inheritance of the seed coat colour in segregating progenies of the interspecific cross between T. alexandrinum and T. apertum. Although, both the parent species possessed yellow seed coat, the F1 seeds were black coloured in the reciprocal cross (T. apertum × T. alexandrinum). Seeds borne on individual F2 plants and the advancing generations segregated in yellow and black seed coat colour, which confirmed xenia effect. F2 seeds collected from individual F1 plants exhibited nine black and seven yellow segregation ratio. The segregation of the seed coat colour recorded from F3 to F5 generations revealed that yellow seed coat was true breeding (i.e. non-segregating) in this interspecific cross (including the reciprocal crosses). However, the black seeded progenies were either true breeding or segregated in nine black: seven yellow ratio or three black: one yellow ratio suggesting a complementary gene interaction or duplicate recessive epistasis. It indicated that the seed coat colour is controlled by complementary gene interaction along with xenia effect in interspecific crosses between T. alexandrinum and T. apertum. Occurrence of the complementary genes across the species could suggest T. apertum to be the progenitor of T. alexandrinum. Inheritance of seed coat colour in reference to its importance in Egyptian clover breeding is also discussed.

  • development characterization and cross species transferability of genomic ssr markers in berseem Trifolium alexandrinum l an important multi cut annual forage legume
    Molecular Breeding, 2015
    Co-Authors: Priyanka Verma, D R Malaviya, P Kaushal, Ajoy Kumar Roy, Amaresh Chandra, Dinesh Pandey, Sabhyata Bhatia
    Abstract:

    Egyptian clover (Trifolium alexandrinum), a multi-cut forage legume, is widely grown as an annual feed crop. Genetic improvement in this legume is largely hampered due to limited availability of genomic resources. Hence, a genomic library enriched for GA/CT motif was constructed to develop SSR primers. As a result, 667 SSR primer pairs were developed and 111 were validated in T. alexandrinum cv. Wardan s-99-1. Among these, 45 primer pairs were used for the analysis of genetic diversity among 86 Trifolium accessions, resulting in 220 alleles with an average of 4.88 alleles per locus. Polymorphic information content for these markers ranged from 0.040 to 0.819 with an average of 0.445. The observed and expected heterozygosity for individual loci ranged from 0.39 to 0.89 and 0.368 to 0.80 with an average of 0.675 and 0.659, respectively. The markers exhibited varied levels of cross-species transferability ranging from 93.33 % across T. resupinatum to 40.0 % across T. balansae. Phylogenetic relationships were analyzed using the unweighted pair group method with arithmetic averages (UPGMA)-based dendrogram which was able to establish the uniqueness of each genotype and grouped them into two major clusters. To the best of our knowledge, the SSR markers reported here are among the first to be reported in this important yet neglected forage legume. They were assessed to be highly efficient and polymorphic and found to be useful for distinguishing the closely related genotypes of Trifolium. This large resource of SSR markers would therefore facilitate new avenues for T. alexandrinum genomics.

  • isozyme variability in Trifolium alexandrinum accessions
    Indian Journal of Genetics and Plant Breeding, 2008
    Co-Authors: B Kumar, D R Malaviya, A K Roy, P Kaushal
    Abstract:

    The present investigation was carried out on 65 accessions of berseem (Trifolium alexandrinum) to estimate the genetic variability based on five enzymes – peroxidase, acid phosphatase, esterase, super oxide dismutase and glutamate oxalo acetate transaminase using starch gel electrophoresis. A similarity matrix showed 95–100% similarity among 64 accessions. Accession EC 329299 belonging to ‘Saidi’ ecotype showed 79% similarity with other accessions. The accessions grouped in three major clusters based on the cutting points in the dendrogram at root level. However, phenetic line drawn at 98% identified 25 groups comprising of 1 to 10 accessions. Of these, 13 groups with more than one germplasm lines showed 100% similarity. Morphologically distinct accessions (i.e. multifoliates and red flowered) formed one cluster. Monomorphic bands (13 out of 26) were in high number. Exotic accessions also showed close similarity to many indigenous accessions which indicated less genetic variability among the germplasm lines introduced from other countries. Polymorphism was recorded for 13 bands (2 for SOD, 3 for GOT, 1 for ACP and 7 for Peroxidase).

  • Trifolium alexandrinum t resupinatum interspecific hybrids developed through embryo rescue
    Plant Cell Tissue and Organ Culture, 2005
    Co-Authors: P Kaushal, B Kumar, D R Malaviya, Ajoy Kumar Roy, A Tiwari
    Abstract:

    This is the first report on the development of interspecific hybrids between T. alexandrinum and T. resupinatum, using embryo rescue. T. resupinatum is an important donor species for traits such as resistance to root and stem rot diseases, and tolerance to alkalinity for the potential improvement of T. alexandrinum. T. alexandrinum, as female parent, was crossed with T. resupinatum and embryos were excised 10–12 days after pollination and cultured in vitro on EC3 medium. Of the recovered embryos, 23.08% showed plumule emergence. For subculturing, shoot-induction medium LSP3 was used. Roots were induced on RL1 medium. Regenerants were hardened and inoculated with Rhizobium before transfer to the field. Morphological appearance of the hybrids was intermediate to both the parents. The hybrids were erect, leafy with short internodes and possessed distinct dwarf bushy compact appearance and were profusely branched. The plants were late flowering with pollen fertility more than 90%. Three out of five hybrid plants set seed following selfing. Hybridity of the plants was also confirmed through Est, PRX, SOD, GOT isozymes and protein banding patterns.

Ajoy Kumar Roy - One of the best experts on this subject based on the ideXlab platform.

  • phenotype study of multifoliolate leaf formation in Trifolium alexandrinum l
    PeerJ, 2021
    Co-Authors: D R Malaviya, P Kaushal, Ajoy Kumar Roy, Shalini Pathak, Ruslan Kalendar
    Abstract:

    Background The genus Trifolium is characterized by typical trifoliolate leaves. Alterations in leaf formats from trifoliolate to multifoliolate, i.e., individual plants bearing trifoliolate, quadrifoliolate, pentafoliolate or more leaflets, were previously reported among many species of the genus. The study is an attempt to develop pure pentafoliolate plants of T. alexandrinum and to understand its genetic control. Methods The experimental material consisted of two populations of T. alexandrinum with multifoliolate leaf expression, i.e.,interspecific hybrid progenies of T. alexandrinum with T. apertum, and T. alexandrinum genotype Penta-1. Penetrance of the multifoliolate trait was observed among multifoliolate and trifoliolate plant progenies. In vitro culture and regeneration of plantlets from the axillary buds from different plant sources was also attempted. Results The inheritance among a large number of plant progenies together with in vitro micro-propagation results did not establish a definite pattern. The multifoliolate leaf formation was of chimeric nature, i.e., more than one leaf format appearing on individual branches. Reversal to normal trifoliolate from multifoliolate was also quite common. Penetrance and expression of multifoliolate leaf formation was higher among the plants raised from multifoliolate plants. Multifoliolate and pure pentafoliolate plants were observed in the progenies of pure trifoliolate plants and vice-versa. There was an apparent increase in the pentafoliolate leaf formation frequency over the years due to targeted selection. A few progenies of the complete pentafoliolate plants in the first year were true breeding in the second year. Frequency of plantlets with multifoliolate leaf formation was also higher in in vitro axillary bud multiplication when the explant bud was excised from the multifoliolate leaf node. Conclusion Number of leaflets being a discrete variable, occurrence of multifoliolate leaves on individual branches, reversal of leaf formats on branches and developing true breeding pentafoliolates were the factors leading to a hypothesis beyond normal Mendelian inheritance. Transposable elements (TEs) involved in leaf development in combination with epigenetics were probably responsible for alterations in the expression of leaflet number. Putative TE's movement owing to chromosomal rearrangements possibly resulted in homozygous pentafoliolate trait with evolutionary significance. The hypothesis provides a new insight into understanding the genetic control of this trait in T. alexandrinum and may also be useful in other Trifolium species where such observations are reported.

  • development characterization and cross species transferability of genomic ssr markers in berseem Trifolium alexandrinum l an important multi cut annual forage legume
    Molecular Breeding, 2015
    Co-Authors: Priyanka Verma, D R Malaviya, P Kaushal, Ajoy Kumar Roy, Amaresh Chandra, Dinesh Pandey, Sabhyata Bhatia
    Abstract:

    Egyptian clover (Trifolium alexandrinum), a multi-cut forage legume, is widely grown as an annual feed crop. Genetic improvement in this legume is largely hampered due to limited availability of genomic resources. Hence, a genomic library enriched for GA/CT motif was constructed to develop SSR primers. As a result, 667 SSR primer pairs were developed and 111 were validated in T. alexandrinum cv. Wardan s-99-1. Among these, 45 primer pairs were used for the analysis of genetic diversity among 86 Trifolium accessions, resulting in 220 alleles with an average of 4.88 alleles per locus. Polymorphic information content for these markers ranged from 0.040 to 0.819 with an average of 0.445. The observed and expected heterozygosity for individual loci ranged from 0.39 to 0.89 and 0.368 to 0.80 with an average of 0.675 and 0.659, respectively. The markers exhibited varied levels of cross-species transferability ranging from 93.33 % across T. resupinatum to 40.0 % across T. balansae. Phylogenetic relationships were analyzed using the unweighted pair group method with arithmetic averages (UPGMA)-based dendrogram which was able to establish the uniqueness of each genotype and grouped them into two major clusters. To the best of our knowledge, the SSR markers reported here are among the first to be reported in this important yet neglected forage legume. They were assessed to be highly efficient and polymorphic and found to be useful for distinguishing the closely related genotypes of Trifolium. This large resource of SSR markers would therefore facilitate new avenues for T. alexandrinum genomics.

  • Trifolium alexandrinum t resupinatum interspecific hybrids developed through embryo rescue
    Plant Cell Tissue and Organ Culture, 2005
    Co-Authors: P Kaushal, B Kumar, D R Malaviya, Ajoy Kumar Roy, A Tiwari
    Abstract:

    This is the first report on the development of interspecific hybrids between T. alexandrinum and T. resupinatum, using embryo rescue. T. resupinatum is an important donor species for traits such as resistance to root and stem rot diseases, and tolerance to alkalinity for the potential improvement of T. alexandrinum. T. alexandrinum, as female parent, was crossed with T. resupinatum and embryos were excised 10–12 days after pollination and cultured in vitro on EC3 medium. Of the recovered embryos, 23.08% showed plumule emergence. For subculturing, shoot-induction medium LSP3 was used. Roots were induced on RL1 medium. Regenerants were hardened and inoculated with Rhizobium before transfer to the field. Morphological appearance of the hybrids was intermediate to both the parents. The hybrids were erect, leafy with short internodes and possessed distinct dwarf bushy compact appearance and were profusely branched. The plants were late flowering with pollen fertility more than 90%. Three out of five hybrid plants set seed following selfing. Hybridity of the plants was also confirmed through Est, PRX, SOD, GOT isozymes and protein banding patterns.

  • affinity between Trifolium alexandrinum and t apertum cytological investigation in embryo rescued hybrid
    Cytologia, 2004
    Co-Authors: D R Malaviya, P Kaushal, Ajoy Kumar Roy, B Kumar
    Abstract:

    Interspecific hybrid of Trifolium alexandrinum L. (Egyptian clover or berseem) with T. apertum Bobrov was developed through embryo rescue. Both the species are annual diploids (2n=16). Chromosomal pairing and fertility of F1 and F2 generation was studied with a view to understand the phylogenetic relationship of the two species and possibility of recombination between their respective genomes. The parental species showed regular bivalent formation followed by normal disjunction, pollen and seed fertility. Among six F1 hybrids, Hybrid 19, 20 and A-15-P1 showed multivalent formations at diakinesis while the rest three showed near normal bivalent formation. Pollen fertility among first generation hybrids was substantially high (>90%) except in Hybrid 19, which showed 78.7% pollen fertility. Variation for pollen size was also observed in Hybrids 19 and 20. 88.75 to 98.25% chromosomes contributed in bivalent formation. Among F2 plants, the meiotic behaviour was near normal and in most of the PMCs, eight bivalent formation was seen. F2 plants B-26-P1 and B-34-P1, however, showed formation of 0.2 quadrivalent and 0.33 trivalent per PMC respectively. Among other F2 plants infrequent formation of univalents was seen. In general, the plants showed more bivalents per PMC than that among F1 plants and 95 to 100% chromosomes associated as bivalents. Pollen fertility among these plants was also more than 95% in majority cases. Fertile F1 hybrids followed by fertile F2 generation and absence of segregation for fertility is an indicator that the two species are not differentiated by sizeable differences in their chromosomal/genic constitution and possesses close affinity.

  • development and characterization of interspecific hybrids of Trifolium alexandrinum t apertum using embryo rescue
    Plant Breeding, 2004
    Co-Authors: D R Malaviya, B Kumar, P Kaushal, Ajoy Kumar Roy, A Tiwari
    Abstract:

    This is the first report on the development of interspecific hybrids between Trifolium alexandrinum and T. apertum using embryo rescue and characterization of F, plants. T. apertum was used as the male parent and T. alexandrinum as the female parent. Development of interspecific hybrids under natural conditions is not successful and so embryo rescue was attempted. Of the several combinations tried, pollination 2 days after emasculation and embryos rescued 11 days after pollination was found to be the best. For embryo culture, EC3 medium consisting of MS basal supplemented with 2.3 μM kinetin and 3% sucrose was used. Germinated embryos were transferred to LSP3 medium 25 days after inoculation wherein most of the cultures showed multiple shoots that were split and subcultured on RL1 medium for rooting. After hardening, about 75% of hybrids were successfully transferred to the field. The hybrids, in general, showed morphological traits intermediate between the two parents; however, a few hybrids showed better growth than either parent. Some F 1 plants were almost 3 weeks later in flowering than the female parent. Pollen fertility among these plants ranged from 78 to approximately 100%. Chromosomal associations at diakinesis and isozyme banding patterns for acid phosphatase (ACP), superoxide dismutase (SOD) and peroxidase also confirmed the hybrid nature of the plants.

A Tiwari - One of the best experts on this subject based on the ideXlab platform.

  • Trifolium alexandrinum t resupinatum interspecific hybrids developed through embryo rescue
    Plant Cell Tissue and Organ Culture, 2005
    Co-Authors: P Kaushal, B Kumar, D R Malaviya, Ajoy Kumar Roy, A Tiwari
    Abstract:

    This is the first report on the development of interspecific hybrids between T. alexandrinum and T. resupinatum, using embryo rescue. T. resupinatum is an important donor species for traits such as resistance to root and stem rot diseases, and tolerance to alkalinity for the potential improvement of T. alexandrinum. T. alexandrinum, as female parent, was crossed with T. resupinatum and embryos were excised 10–12 days after pollination and cultured in vitro on EC3 medium. Of the recovered embryos, 23.08% showed plumule emergence. For subculturing, shoot-induction medium LSP3 was used. Roots were induced on RL1 medium. Regenerants were hardened and inoculated with Rhizobium before transfer to the field. Morphological appearance of the hybrids was intermediate to both the parents. The hybrids were erect, leafy with short internodes and possessed distinct dwarf bushy compact appearance and were profusely branched. The plants were late flowering with pollen fertility more than 90%. Three out of five hybrid plants set seed following selfing. Hybridity of the plants was also confirmed through Est, PRX, SOD, GOT isozymes and protein banding patterns.

  • development and characterization of interspecific hybrids of Trifolium alexandrinum t apertum using embryo rescue
    Plant Breeding, 2004
    Co-Authors: D R Malaviya, B Kumar, P Kaushal, Ajoy Kumar Roy, A Tiwari
    Abstract:

    This is the first report on the development of interspecific hybrids between Trifolium alexandrinum and T. apertum using embryo rescue and characterization of F, plants. T. apertum was used as the male parent and T. alexandrinum as the female parent. Development of interspecific hybrids under natural conditions is not successful and so embryo rescue was attempted. Of the several combinations tried, pollination 2 days after emasculation and embryos rescued 11 days after pollination was found to be the best. For embryo culture, EC3 medium consisting of MS basal supplemented with 2.3 μM kinetin and 3% sucrose was used. Germinated embryos were transferred to LSP3 medium 25 days after inoculation wherein most of the cultures showed multiple shoots that were split and subcultured on RL1 medium for rooting. After hardening, about 75% of hybrids were successfully transferred to the field. The hybrids, in general, showed morphological traits intermediate between the two parents; however, a few hybrids showed better growth than either parent. Some F 1 plants were almost 3 weeks later in flowering than the female parent. Pollen fertility among these plants ranged from 78 to approximately 100%. Chromosomal associations at diakinesis and isozyme banding patterns for acid phosphatase (ACP), superoxide dismutase (SOD) and peroxidase also confirmed the hybrid nature of the plants.

  • interspecific hybridization of Trifolium alexandrinum with t constantinopolitanum using embryo rescue
    Plant Cell Reports, 2004
    Co-Authors: Ajoy Kumar Roy, B Kumar, D R Malaviya, P Kaushal, A Tiwari
    Abstract:

    The embryo rescue technique was successfully used to raise hybrids between Trifolium alexandrinum and T. constantinopolitanum. As a result of its narrow genetic base, genetic improvement in Egyptian clover (syn. Berseem; T. alexandrinum), an important fodder crop in tropical and subtropical countries, is hampered, thereby making it imperative to introduce alien genes from related species. In a conventional interspecific hybridization program, hybrids could not be raised due to post-fertilization barriers. Of the several combinations tried, pollination 2 days after emasculation was found to be the best. Globular embryos were observed 5–7 days after pollination (DAP), followed by heart-shaped embryos 10–12 DAP. Embryos excised at the heart-shaped stage responded well to EC3 culture medium. Of 612 crosses, 33 healthy embryos could be excised and cultured on EC3 medium. The plumule emerged 8–12 days following inoculation. The embryo-rescued plants were hardened, inoculated with Rhizobium and transferred to the field. The hybrids showed intermediate morphological features with reduced pollen fertility (55–65%) and a chromosomal complement of 2n=16. Biochemical characterization using isozymes confirmed hybridity.

B Kumar - One of the best experts on this subject based on the ideXlab platform.

  • isozyme variability in Trifolium alexandrinum accessions
    Indian Journal of Genetics and Plant Breeding, 2008
    Co-Authors: B Kumar, D R Malaviya, A K Roy, P Kaushal
    Abstract:

    The present investigation was carried out on 65 accessions of berseem (Trifolium alexandrinum) to estimate the genetic variability based on five enzymes – peroxidase, acid phosphatase, esterase, super oxide dismutase and glutamate oxalo acetate transaminase using starch gel electrophoresis. A similarity matrix showed 95–100% similarity among 64 accessions. Accession EC 329299 belonging to ‘Saidi’ ecotype showed 79% similarity with other accessions. The accessions grouped in three major clusters based on the cutting points in the dendrogram at root level. However, phenetic line drawn at 98% identified 25 groups comprising of 1 to 10 accessions. Of these, 13 groups with more than one germplasm lines showed 100% similarity. Morphologically distinct accessions (i.e. multifoliates and red flowered) formed one cluster. Monomorphic bands (13 out of 26) were in high number. Exotic accessions also showed close similarity to many indigenous accessions which indicated less genetic variability among the germplasm lines introduced from other countries. Polymorphism was recorded for 13 bands (2 for SOD, 3 for GOT, 1 for ACP and 7 for Peroxidase).

  • Trifolium alexandrinum t resupinatum interspecific hybrids developed through embryo rescue
    Plant Cell Tissue and Organ Culture, 2005
    Co-Authors: P Kaushal, B Kumar, D R Malaviya, Ajoy Kumar Roy, A Tiwari
    Abstract:

    This is the first report on the development of interspecific hybrids between T. alexandrinum and T. resupinatum, using embryo rescue. T. resupinatum is an important donor species for traits such as resistance to root and stem rot diseases, and tolerance to alkalinity for the potential improvement of T. alexandrinum. T. alexandrinum, as female parent, was crossed with T. resupinatum and embryos were excised 10–12 days after pollination and cultured in vitro on EC3 medium. Of the recovered embryos, 23.08% showed plumule emergence. For subculturing, shoot-induction medium LSP3 was used. Roots were induced on RL1 medium. Regenerants were hardened and inoculated with Rhizobium before transfer to the field. Morphological appearance of the hybrids was intermediate to both the parents. The hybrids were erect, leafy with short internodes and possessed distinct dwarf bushy compact appearance and were profusely branched. The plants were late flowering with pollen fertility more than 90%. Three out of five hybrid plants set seed following selfing. Hybridity of the plants was also confirmed through Est, PRX, SOD, GOT isozymes and protein banding patterns.

  • affinity between Trifolium alexandrinum and t apertum cytological investigation in embryo rescued hybrid
    Cytologia, 2004
    Co-Authors: D R Malaviya, P Kaushal, Ajoy Kumar Roy, B Kumar
    Abstract:

    Interspecific hybrid of Trifolium alexandrinum L. (Egyptian clover or berseem) with T. apertum Bobrov was developed through embryo rescue. Both the species are annual diploids (2n=16). Chromosomal pairing and fertility of F1 and F2 generation was studied with a view to understand the phylogenetic relationship of the two species and possibility of recombination between their respective genomes. The parental species showed regular bivalent formation followed by normal disjunction, pollen and seed fertility. Among six F1 hybrids, Hybrid 19, 20 and A-15-P1 showed multivalent formations at diakinesis while the rest three showed near normal bivalent formation. Pollen fertility among first generation hybrids was substantially high (>90%) except in Hybrid 19, which showed 78.7% pollen fertility. Variation for pollen size was also observed in Hybrids 19 and 20. 88.75 to 98.25% chromosomes contributed in bivalent formation. Among F2 plants, the meiotic behaviour was near normal and in most of the PMCs, eight bivalent formation was seen. F2 plants B-26-P1 and B-34-P1, however, showed formation of 0.2 quadrivalent and 0.33 trivalent per PMC respectively. Among other F2 plants infrequent formation of univalents was seen. In general, the plants showed more bivalents per PMC than that among F1 plants and 95 to 100% chromosomes associated as bivalents. Pollen fertility among these plants was also more than 95% in majority cases. Fertile F1 hybrids followed by fertile F2 generation and absence of segregation for fertility is an indicator that the two species are not differentiated by sizeable differences in their chromosomal/genic constitution and possesses close affinity.

  • development and characterization of interspecific hybrids of Trifolium alexandrinum t apertum using embryo rescue
    Plant Breeding, 2004
    Co-Authors: D R Malaviya, B Kumar, P Kaushal, Ajoy Kumar Roy, A Tiwari
    Abstract:

    This is the first report on the development of interspecific hybrids between Trifolium alexandrinum and T. apertum using embryo rescue and characterization of F, plants. T. apertum was used as the male parent and T. alexandrinum as the female parent. Development of interspecific hybrids under natural conditions is not successful and so embryo rescue was attempted. Of the several combinations tried, pollination 2 days after emasculation and embryos rescued 11 days after pollination was found to be the best. For embryo culture, EC3 medium consisting of MS basal supplemented with 2.3 μM kinetin and 3% sucrose was used. Germinated embryos were transferred to LSP3 medium 25 days after inoculation wherein most of the cultures showed multiple shoots that were split and subcultured on RL1 medium for rooting. After hardening, about 75% of hybrids were successfully transferred to the field. The hybrids, in general, showed morphological traits intermediate between the two parents; however, a few hybrids showed better growth than either parent. Some F 1 plants were almost 3 weeks later in flowering than the female parent. Pollen fertility among these plants ranged from 78 to approximately 100%. Chromosomal associations at diakinesis and isozyme banding patterns for acid phosphatase (ACP), superoxide dismutase (SOD) and peroxidase also confirmed the hybrid nature of the plants.

  • interspecific hybridization of Trifolium alexandrinum with t constantinopolitanum using embryo rescue
    Plant Cell Reports, 2004
    Co-Authors: Ajoy Kumar Roy, B Kumar, D R Malaviya, P Kaushal, A Tiwari
    Abstract:

    The embryo rescue technique was successfully used to raise hybrids between Trifolium alexandrinum and T. constantinopolitanum. As a result of its narrow genetic base, genetic improvement in Egyptian clover (syn. Berseem; T. alexandrinum), an important fodder crop in tropical and subtropical countries, is hampered, thereby making it imperative to introduce alien genes from related species. In a conventional interspecific hybridization program, hybrids could not be raised due to post-fertilization barriers. Of the several combinations tried, pollination 2 days after emasculation was found to be the best. Globular embryos were observed 5–7 days after pollination (DAP), followed by heart-shaped embryos 10–12 DAP. Embryos excised at the heart-shaped stage responded well to EC3 culture medium. Of 612 crosses, 33 healthy embryos could be excised and cultured on EC3 medium. The plumule emerged 8–12 days following inoculation. The embryo-rescued plants were hardened, inoculated with Rhizobium and transferred to the field. The hybrids showed intermediate morphological features with reduced pollen fertility (55–65%) and a chromosomal complement of 2n=16. Biochemical characterization using isozymes confirmed hybridity.

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