Finger Millet Genotypes Research Articles

Tracing QTLs for Leaf Blast Resistance and Agronomic Performance of Finger Millet (Eleusine coracana (L.) Gaertn.) Genotypes through Association Mapping and in silico Comparative Genomics Analyses.

Finger millet is one of the small millets with high nutritive value. This crop is vulnerable to blast disease caused by Pyricularia grisea, which occurs annually during rainy and winter seasons. Leaf blast occurs at early crop stage and is highly damaging. Mapping of resistance genes and other quantitative trait loci (QTLs) for agronomic performance can be of great use for improving finger millet genotypes. Evaluation of one hundred and twenty-eight finger millet genotypes in natural field conditions revealed that leaf blast caused severe setback on agronomic performance for susceptible genotypes, most significant traits being plant height and root length. Plant height was reduced under disease severity while root length was increased. Among the genotypes, IE4795 showed superior response in terms of both disease resistance and better agronomic performance. A total of seven unambiguous QTLs were found to be associated with various agronomic traits including leaf blast resistance by association mapping analysis. The markers, UGEP101 and UGEP95, were strongly associated with blast resistance. UGEP98 was associated with tiller number and UGEP9 was associated with root length and seed yield. Cross species validation of markers revealed that 12 candidate genes were associated with 8 QTLs in the genomes of grass species such as rice, foxtail millet, maize, Brachypodium stacei, B. distachyon, Panicum hallii and switchgrass. Several candidate genes were found proximal to orthologous sequences of the identified QTLs such as 1,4-β-glucanase for leaf blast resistance, cytokinin dehydrogenase (CKX) for tiller production, calmodulin (CaM) binding protein for seed yield and pectin methylesterase inhibitor (PMEI) for root growth and development. Most of these QTLs and their putatively associated candidate genes are reported for first time in finger millet. On validation, these novel QTLs may be utilized in future for marker assisted breeding for the development of fungal resistant and high yielding varieties of finger millet.

Over-expression of a NAC 67 transcription factor from finger millet (Eleusine coracana L.) confers tolerance against salinity and drought stress in rice.

BackgroundNAC proteins (NAM (No apical meristem), ATAF (Arabidopsis transcription activation factor) and CUC (cup-shaped cotyledon)) are plant-specific transcription factors reported to be involved in regulating growth, development and stress responses. Salinity responsive transcriptome profiling in a set of contrasting finger millet genotypes through RNA-sequencing resulted in the identification of a NAC homolog (EcNAC 67) exhibiting differential salinity responsive expression pattern.MethodsFull length cDNA of EcNAC67 was isolated, characterized and validated for its role in abiotic stress tolerance through agrobacterium mediated genetic transformation in a rice cultivar ASD16.ResultsBioinformatics analysis of putative NAC transcription factor (TF) isolated from a salinity tolerant finger millet showed its genetic relatedness to NAC67 family TFs in related cereals. Putative transgenic lines of rice over-expressing EcNAC67 were generated through Agrobacterium mediated transformation and presence/integration of transgene was confirmed through PCR and southern hybridization analysis. Transgenic rice plants harboring EcNAC67 showed enhanced tolerance against drought and salinity under greenhouse conditions. Transgenic rice plants were found to possess higher root and shoot biomass during stress and showed better revival ability upon relief from salinity stress. Upon drought stress, transgenic lines were found to maintain higher relative water content and lesser reduction in grain yield when compared to non-transgenic ASD16 plants. Drought induced spikelet sterility was found to be much lower in the transgenic lines than the non-transgenic ASD16.ConclusionResults revealed the significant role of EcNAC67 in modulating responses against dehydration stress in rice. No detectable abnormalities in the phenotypic traits were observed in the transgenic plants under normal growth conditions. Results indicate that EcNAC67 can be used as a novel source for engineering tolerance against drought and salinity stress in rice and other crop plants.Electronic supplementary materialThe online version of this article (doi:10.1186/s12896-016-0261-1) contains supplementary material, which is available to authorized users.

Genetic polymorphisms among and between blast disease resistant and susceptible finger millet,Eleusine coracana(L.) Gaertn.

AbstractFungal blast disease is one of the major constraints in finger millet production. Breeding for disease resistance in finger millet, needs characterization of genetic polymorphism among and between the resistant and susceptible genotypes. In total, 67 finger millet genotypes, which are resistant or susceptible to fungal blast disease, were analysed using sequence-related amplified polymorphism (SRAP) and simple sequence repeat (SSR) markers to assess genetic variations and select diverse parents. Twelve each of SRAP and SSR primers produced 95.1 and 93.1% polymorphic bands and grouped them into unweighted pair-group method with arithmetic average clusters. Two of the finger millet genotypes, IE 4709 (blast resistant) and INDAF 7 (susceptible) were distinguished as most diverse genotypes as parents. Several genotype-specific bands observed with SSR primers are potential in developing genotype-specific markers. A high genetic diversity within the resistant and susceptible genotypes, rather than between them, was revealed through Nei's gene diversity (h) index and analysis of molecular variance. The finding helps us to understand the extent of genetic polymorphism between blast disease resistant and susceptible finger millet genotypes to exploit in resistance breeding programs.

Genetic diversity for grain Zn concentration in finger millet genotypes: Potential for improving human Zn nutrition

Nearly half of the world population suffers from micronutrient malnutrition, particularly Zn deficiency. It is important to understand genetic variation for uptake and translocation behaviors of Zn in relevant crop species to increase Zn concentration in edible parts. In the present study, genetic variation in grain Zn concentration of 319 finger millet genotypes was assessed. Large genetic variation was found among the genotypes, with concentrations ranging from 10 to 86μgg−1 grain. Uptake and translocation studies with Zn/65Zn application in 12 selected low-Zn genotypes showed wide variation in root uptake and shoot translocation, with genotypes GEC331 and GEC164 showing greater uptake and translocation. Genotypes GEC164 and GEC543 showed increased grain Zn concentration. Genotypes GEC331 and GEC164 also showed improved yield under Zn treatment. Appreciable variation in grain Zn concentration among finger millet genotypes found in this study offers opportunities to improve Zn nutrition through breeding.

Assessment of genetic diversity, population structure and relationships in Indian and non-Indian genotypes of finger millet (Eleusine coracana (L.) Gaertn) using genomic SSR markers.

We evaluated the genetic variation and population structure in Indian and non-Indian genotypes of finger millet using 87 genomic SSR primers. The 128 finger millet genotypes were collected and genomic DNA was isolated. Eighty-seven genomic SSR primers with 60–70 % GC contents were used for PCR analysis of 128 finger millet genotypes. The PCR products were separated and visualized on a 6 % polyacrylamide gel followed by silver staining. The data were used to estimate major allele frequency using Power Marker v3.0. Dendrograms were constructed based on the Jaccard’s similarity coefficient. Statistical fitness and population structure analyses were performed to find the genetic diversity. The mean major allele frequency was 0.92; the means of polymorphic alleles were 2.13 per primer and 1.45 per genotype; the average polymorphism was 59.94 % per primer and average PIC value was 0.44 per primer. Indian genotypes produced an additional 0.21 allele than non-Indian genotypes. Gene diversity was in the range from 0.02 to 0.35. The average heterozygosity was 0.11, close to 100 % homozygosity. The highest inbreeding coefficient was observed with SSR marker UGEP67. The Jaccard’s similarity coefficient value ranged from 0.011 to 0.836. The highest similarity value was 0.836 between genotypes DPI009-04 and GPU-45. Indian genotypes were placed in Eleusine coracana major cluster (EcMC) 1 along with 6 non-Indian genotypes. AMOVA showed that molecular variance in genotypes from various geographical regions was 4 %; among populations it was 3 % and within populations it was 93 %. PCA scatter plot analysis showed that GPU-28, GPU-45 and DPI009-04 were closely dispersed in first component axis. In structural analysis, the genotypes were divided into three subpopulations (SP1, SP2 and SP3). All the three subpopulations had an admixture of alleles and no pure line was observed. These analyses confirmed that all the genotypes were genetically diverse and had been grouped based on their geographic regions.

Effect of Zn application on its uptake, distribution and concentration of Fe and Cu in finger millet [Eleusine coracana (L.) Gaertn.

ABSTRACTZinc (Zn) malnutrition can be alleviated by increasing the dietary Zn intake through Zn biofortification of edible crops. Agronomic and genetic biofortification has been suggested as better option to increase the dietary Zn. In this study, we show considerable genetic variability for seed Zn concentration in six leading finger millet genotypes. External application of Zn resulted in improved Zn concentration in different plant parts; in particular there was significant increase in seed Zn concentration in all genotypes. Though genotypes GPU28 and INDAF5 showed differences in root and shoot Zn at vegetative stage but at reproductive stage there was no significant difference. Apart from that, Zn application increased the seed iron (Fe) concentration with no or minimal effect on copper (Cu) concentration.

Development and evaluation of early maturing white-grained finger millet (Eleusine coracana L.) genotypes for cultivation in sub-mountain Himalayan region of India

White grained finger millet genotypes has become a thrust area in finger millet breeding due to increased demand of non-glutinous food products and lesser acceptability of brown grained finger millet. Sixteen white grain finger millet lines were developed by crossing extra early maturing brown grained finger millet genotypes including adapted varieties with late maturing white grained finger millet genotypes. The quantitative data of 16 lines along with parents were subjected to multivariate analysis. A wide range of variation was observed for all the studied traits. The parental lines of brown and white grained finger millet genotypes exhibited extreme values for grain yield and days to maturity whereas, the developed white grained genotypes showed moderate values. Projections of genotypes in PCA biplot showed close association of newly developed white grain genotypes VL 427, VL 360, VR 485, VR 443, VL 366, VR 425B, VR 425A and VL 356 with VL 201 (brown type) because of earliness and high yield potential. The cluster analysis further indicated that white grained genotypes from second cluster are probable candidates for further testing and release, and further refinement in breeding strategy by hybridizing white genotypes from second cluster with brown genotype in third cluster for incorporating earliness and high yield.

Evaluation of finger millet genotypes for partial resistance to leaf blast caused by Pyricularia grisea

Blast of finger millet (ragi) caused by Pyricularia grisea is disastrous disease to limit the potential yield levels in the farmers field. The pathogen affects the crop at all growth stages and the most susceptible stage for leaf blast is seedling stage. Use of resistant cultivars for minimizing losses due to leaf blast at early stage of crop growth is a viable and economical alternative. In the present study, partial resistance to leaf blast in fourteen genotypes of finger millet was assessed under field conditions. None of the genotypes were highly resistant to leaf blast. However, two genotypes namely PR 10-30 and VL 368 were shown resistance. Seven genotypes namely VR 988, VL 353, VL 348, TNAU 1214, GPU 45, VL 369, KOPM 942 were found moderately resistant, three namely PPR 2773, KRI 007-01, VR 708 were susceptible and two namely TNAU 1226 and REC 69 were highly susceptible. The correlation coefficients between leaf blast severities (LBS) with different components of partial resistance of slow blasting were significant. Low value of coefficient of infection (CI), apparent infection rate (r), area under disease progress curve (AUDPC), lesion number (LN), lesion area (LA), lesion circumference (LC) and high value of incubation period (IP) were recorded in resistant genotypes of finger millet, whereas, higher values of CI, r, AUDPC, LN, LA, LC and low values of IP were recorded in highly susceptible genotypes. A significant positive correlation was found in leaf blast severity with CI (r=0.948*), r (r=0.829*), AUDPC (r=0.998*), LN (r=0.967*), LA (r=0.957*), LC (r=0.976*) whereas IP (r=-0.825*) was negatively correlated with blast resistance. PR 10-30 and VL 368 showed higher degree of partial resistance or slow blasting resistance against P. grisea infection on finger millet.

Stability of finger millet genotypes under diverse environments

The present study aimed at deciphering the stability and patterns of genotype × environment interaction (GEI) in finger millet genotypes tested in All India Coordinated Trials using GGE biplot technique. The combined ANOVA for grain yield of five finger millet cultivars at four environments showed that environments (E), genotypes (G) and GEI were highly significant (P<0.01). GGE biplot grouped the four environments into two mega environments with GPU 67 and VR 990 as winning genotypes. VL 376 was found to be an ideal genotype in terms of high yield and stability followed by VR 990 as desirable genotype.

Evaluation of finger millet genotypes for stability using parametric and non-parametric methods in India

A study of phenotypic stability of 30 finger millet genotypes was conducted to assess genotype-environment interaction (GEI) and identify stable finger millet ( Eleusine coracana (L.) Gaertn. subsp. coracana ) genotypes for grain yield across four diverse locations in India. Genotypes, environments main effects and GEI were significant at P < 0.01. Different stability measures identified different wide adaptable genotypes. The parameters Wi 2 , σi 2 , Si (1) and Si (2) identified TNEC 1234, GPU 67, KMR 344, DHFMV 10-2-1, GPU 92 and VL 352 as stable genotypes, similarly ASV identified PPR 1044, PPR 1040, BR 45, GPU 91, VL 384 and DHFMV 78-3-1 while GPU 67, VL 376, DHFMV 10-2-1, GPU 92, VR 990 and GPU 91 were top six stable genotypes according to Ysi. The rank correlation matrix identified only two important stability measures (Ysi and YSI) in the identification of stable genotypes without compromise for grain yield. Among these two, Ysi was found to be the better choice for screening of genotypes for both yield and stability. Keywords G×E, Parametric stability, Non parametric stability, Rank Correlation, Finger millet

AMMI Analysis of G X E Interaction in Finger Millet Genotypes for Yield Stability and Adaptability

Multilocation yield trials on fifteen early duration finger millet (Eleusine coracana Gaertn.) genotypes were conducted under early and late transplanting condition at Bhubaneswar and early direct sown condition at Berhmpur for three years during kharif season. G X E interaction analysis of grain yield in AMMI (Additive Main and Multiplicative Interaction) model showed differential interaction of the genotypes in three planting conditions. IPCA-1 explained 66.5% of G X E interaction. AMMI I biplot of G X E interaction showed that RAU 8, DM 7, SRS 2 and BM 107-2 had high mean and moderate negative interaction effects, while Bhairabi and Neelachal had high mean and small positive interaction effects. AMMI biplot showed differential interaction of the genotypes in different environmental conditions. On the basis of AMMI II predicted yield, the genotypes SRS 2 and RAU 8 would be better adapted to all planting conditions. Genotype Bhairabi would be suited for early cropping, but not late cropping. The genotype DM 7 would be best suited for transplanting condition and not for the direct seeded condition. BM 107-2 would be best suited for late transplanting, while Neelachal would be best suited for the direct seeded condition, indicating their specific adaptation.

Population Structure and Genetic Variation in Indian and African Eleusine coracana (L.) Gaertn

Population structure and genetic variation was estimated for 40 Indian and African finger millet genotypes using 237 random amplified polymorphic DNA (RAPD) and 16 morphological markers revealing differences between African and Indian groups. African genotypes showed 53.59%; 0.17±0.19; 0.27±0.27 compared to 48.10%; 0.15±0.19; 0.24±0.27 polymorphism, Nei's gene diversity and Shannon's Information index, respectively in Indian genotypes. Primers OPM-20, OPW-04, OPX-11 were identified as the most informative primers in terms of high % polymorphism and Nei's gene diversity values. Population structure analysis revealed the existence of two distinct sub-groups, correlated well with primary and secondary centers of finger millet diversity and can be utilized to enhance and widen finger millet germplasm base.

Response of Elite Kenyan Finger Millet (&lt;i&gt;Eleusine coracana, &lt;/i&gt;L. Gaertn) Genotypes to Ethrel Application

Finger millet is a staple food crop of many communities in Africa. The crop is highly nutritious and has incredible grain storage quality. Limited research investment in finger millet in the past has resulted in poor yields and there are currently no commercial hybrids. We investigated the response of different finger millet genotypes (Okhale-1, Gulu-E, KACCIMMI-72, IE 2872, IE 4115 and U-15) to the application of a plant growth regulator hormone (Ethrel). Six elite Kenyan finger millet varieties with contrasting agronomic traits were crossed in a 6 x 6 diallel pattern. To enhance male sterility across female parents, we subjected the plants to Ethrel at concentrations of 1,500ppm, 1,750ppm and 2,000ppm against a 0ppm check. Dwarfing of sprayed plants that resulted in less lodging and ultimately higher yields were observed among plants sprayed with Ethrel at different concentrations. Ethrel application at 2,000ppm had the most dwarfing effect on plants while spraying plants with 1,500ppm of Ethrel resulted in increased grain weight. Although our results demonstrate overall positive effect of Ethrel on finger millet production, the optimum concentrations for more efficient hybridization will still need to be determined.

Response of Elite Kenyan Finger Millet (&lt;i&gt;Eleusine coracana, &lt;/i&gt;L. Gaertn) Genotypes to Ethrel Application

Finger millet is a staple food crop of many communities in Africa. The crop is highly nutritious and has incredible grain storage quality. Limited research investment in finger millet in the past has resulted in poor yields and there are currently no commercial hybrids. We investigated the response of different finger millet genotypes (Okhale-1, Gulu-E, KACCIMMI-72, IE 2872, IE 4115 and U-15) to the application of a plant growth regulator hormone (Ethrel). Six elite Kenyan finger millet varieties with contrasting agronomic traits were crossed in a 6 x 6 diallel pattern. To enhance male sterility across female parents, we subjected the plants to Ethrel at concentrations of 1,500ppm, 1,750ppm and 2,000ppm against a 0ppm check. Dwarfing of sprayed plants that resulted in less lodging and ultimately higher yields were observed among plants sprayed with Ethrel at different concentrations. Ethrel application at 2,000ppm had the most dwarfing effect on plants while spraying plants with 1,500ppm of Ethrel resulted in increased grain weight. Although our results demonstrate overall positive effect of Ethrel on finger millet production, the optimum concentrations for more efficient hybridization will still need to be determined.

In vitro plant regeneration from mature seeds of finger millet (Eleusine coracana) through somatic embryogenesis

Mature seeds of two genotypes of finger millet, viz., PR 202 and PES 400, were cultured on 20 different callus induction media containing four levels of 2,4-d (1, 1.5, 2.0 and 2.5 mg l−1) alone or in combination with two levels of cytokinin (0.5 and 1.0 mg l−1 kinetin or BAP). 2,4-d was found to be most effective for callus induction and development when combined with kinetin. Genotypic response for embryogenic callus development ranged from 29.5 to 78.2 % for PR 202 and 10 to 35.7 % for PES 400. After 45–50 days of culture, calli were transferred in light to develop into green nodular calli. Plantlets were obtained by transferring the green nodular embryogenic calli onto regeneration media containing different concentrations and combinations of GA3, kinetin and BAP. Kinetin was found to be more effective for shoot regeneration as compared to BAP. When Callus derived from BAP containing media were transferred on regeneration media containing GA3 alone or with BAP, developed shoots were either devoid of roots or had few thin roots. These shoots were rooted on MS basal media and MS media supplemented with IBA (0.5 or 1.0 mg l−1), NAA (1 mg l−1) and IBA + NAA (0.5 mg l−1 each). Rooting media containing 0.5 mg l−1 IBA and 0.5 mg l−1 NAA showed remarkable rooting response, inducing a thick bunch of 11.5–12 healthy, white and long roots per shoot. Regenerated plants were transferred to pots containing sterilized soil:sand:vermiculite:FYM (2:2:2:1) and were irrigated with Hoagland solution. Regenerated plants grew well and were successfully established in soil.

Influence of plant growth regulators and spermidine on somatic embryogenesis and plant regeneration in four Indian genotypes of finger millet (Eleusine coracana (L.) Gaertn)

Effect of plant growth regulators and spermidine on somatic embryogenesis and regeneration was investigated in finger millet. Mature embryos, and 3 days old seedling-derived shoot apical meristems were cultured on Murashige and Skoog (MS) medium containing picloram, 2,4,5-trichlorophenoxyacetic acid, 2,4-dichlorophenoxyacetic acid (2,4-D) and 1-naphthaleneacetic acid. Improved embryogenesis (84.7 %) was found on MS medium containing 4.0 mg L−1 2,4-D and 0.5 mg L−1 kinetin in both explants. MS medium containing 1.5 mM spermidine along with 4.0 mg L−1 2,4-D and 0.5 mg L−1 kinetin produced highest frequency (90.4 %) of somatic embryogenesis from mature embryo derived callus in genotype ‘CO(Ra)-14’. On same medium somatic embryogenesis frequencies of ‘GPU-25’, ‘Try-1’ and ‘Piyur-2’ genotypes were 55.5, 85.3 and 58.7 %, respectively after 4 weeks of incubation in dark. MS medium containing 4.0 mg L−1 6-benzylaminopurine, 0.2 mg L−1 2,4-D and 1.5 mM spermidine was found to be optimum for shoot regeneration or somatic embryos in all four genotypes of finger millet. We also studied the influence of exogenous spermidine (2.0–4.0 mM) on regeneration of ‘CO(Ra)-14’ mature embryo-derived new and long-term (20–180 days old) calluses. Highest regeneration frequency 93.1 % and mean number of shoots 25.5 were produced on MS medium containing 3.0 mM spermidine, 4.0 mg L−1 BAP and 0.2 mg L−1 2,4-D using 60 days old callus. Regenerated shoots effectively rooted on half-strength MS medium and successfully acclimatized in soil with 100 % survival rate and they grew normally without showing any morphological variation. Genetic variation of in vitro derived plants and control plants were analyzed by RAPD markers.

Using molecular markers to assess the genetic diversity and population structure of finger millet (Eleusine coracana (L.) Gaertn.) from various geographical regions

A genetic diversity study of 128 finger millet genotypes collected from various geographical regions was performed using RAPD markers. The average locus obtained across all genotypes was 115.56 per primer and 0.9 per genotype; the average polymorphism was 76.48 % per primer and average PIC value was 0.40 per primer. The Jaccard’s similarity coefficient value ranged from 0.0085 to 0.81, and UPGMA cluster analysis showed that the bootstrap value was 100 %. These analyses confirmed that all the genotypes were genetically diverse. The genotypes have been grouped into twelve subclusters. Paiyur-2 and KRI007-01 showed the highest Jaccard’s similarity value of 0.81 in UPGMA analysis. All Indian genotypes were placed in subclusters EcC1 to EcC7 along with nine non-Indian genotypes based on UPGMA analysis. AMOVA analysis showed that the percentage of molecular variance among the various geographical regions was 1 %, among populations it was 5 % and within populations it was 94 %. PCA analysis revealed that first and third component axes accounted for 11.3 and 3.7 % of total variance respectively and genotypes were distributed according to their various geographical regions. The correlation value ranged from −0.4 to 0.5 in PCA loadings analysis. Cophenetic correlation coefficient value was 0.8802. In structural analysis, the genotypes were divided into four subpopulations (SP1, SP2, SP3 and SP4) and it revealed that all the four subpopulations had an admixture of alleles and only one pure line (Paiyur-2) was observed. There was good correspondence between the Radial tree analysis and the population structure. This study may form the basis for finger millet breeding and improvement program.

Identification and characterization of calcium transporter gene family in finger millet in relation to grain calcium content

Calcium (Ca) is an essential mineral for proper growth and development of plants as well as animals. In plants including cereals, calcium is deposited in seed during its development which is mediated by specialized Ca transporters. Common cereal seeds contain very low amounts of Ca while the finger millet (Eleusine coracana) contains exceptionally high amounts of Ca in seed. In order to understand the role of Ca transporters in grain Ca accumulation, developing seed transcriptome of two finger millet genotypes (GP-1, low Ca and GP-45 high Ca) differing in seed Ca content was sequenced using Illumina paired-end sequencing technology and members of Ca transporter gene family were identified. Out of 109,218 and 120,130 contigs, 86 and 81 contigs encoding Ca transporters were identified in GP-1 and GP-45, respectively. After removal of redundant sequences, a total of 19 sequences were confirmed as Ca transporter genes, which includes 11 Ca2+ ATPases, 07 Ca2+/cation exchangers and 01 Ca2+ channel. The differential expressions of all genes were analyzed from transcriptome data and it was observed that 9 and 3 genes were highly expressed in GP-45 and GP-1 genotypes respectively. Validation of transcriptome expression data of selected Ca transporter genes was performed on different stages of developing spikes of both genotypes grown under different concentrations of exogenous Ca. In both genotypes, significant correlation was observed between the expression of these genes, especially EcCaX3, and on the amount of Ca accumulated in seed. The positive correlation of seed mass with the amount of Ca concentration was also observed. The efficient Ca transport property and responsiveness of EcCAX3 towards exogenous Ca could be utilized in future biofortification program.

Direct plant regeneration from in vitro-derived shoot apical meristems of finger millet (Eleusine coracana (L.) Gaertn.)

A rapid and reproducible protocol for direct plant regeneration has been established using in vitro-derived, actively growing shoot apical meristems (SAMs) of the genotype 'CO(Ra)-14'. Shoot apical meristems from 3-day-old seedlings of three finger millet genotypes viz, 'CO(Ra)-14', 'Try-1', and 'Paiyur-2' were assessed for the efficiency of direct shoot induction. The regeneration efficiency of 'CO(Ra)-14' surpassed the other two genotypes and produced a mass of green, multiple shoots within 12 d on Murashige and Skoog medium containing 17.6 μM 6-benzylaminopurine (BA), 0.9 μM 2,4-dichlorophenoxyacetic acid in combination with 750 mg L−1 proline, 500 mg L−1 casein enzymatic hydrolysate, and 2 mg L−1 glycine. 'CO(Ra)-14' showed a maximum frequency of multiple shoot regeneration (96%) with an average of 8.3 shoots per explant. The age and size of shoot apical meristems played a crucial role in triggering adventitious shoot bud proliferation. The multiple shoot buds elongated spontaneously upon reducing the concentration of BA (to 11.0 μM) in the shoot induction medium. The in vitro proliferated shoots were rooted best in half-strength MS medium containing 2.8 μM indole-3-acetic acid and successfully acclimated in the field, subsequently developing into fertile plants. Thus, we report a short tenure (45 d) in vitro whole plant regeneration system, which could be effectively utilized for the production of transgenic finger millet plants.

Analysis of Genetic Variations inEleusine coracana(L.) genotypes using ISSR Molecular Markers

Inter Simple Sequence Repeat (ISSR) markers were used to assess the genetic rela-tionships among 27 finger millet genotypes. Ten 5 ’anchored and ten 3 ’anchored ISSR primers were initially screened for PCR amplifications. From a total of 82 DNA fragments produced, 76 (92.68%) were found to be polymorphic. The number of bands produced by different markers was ranging from 2 to 8. PIC value and Rp value were ranging from 0.143 to 0.983 and 0.52 to 11.56, respectively. The genetic similarity matrix varied from 0.43 to 0.98 in these genotypes. Dendogram analysis revealed formation of two major groups, one with five subgroups compris-ing 10 genotypes and the other one consisted of 17 genotypes in five subgroups.. The results re-vealed that ISSR markers could be efficiently used to quickly access the genetic variation available in the finger millet genotypes.