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Not AvailableAntioxidant enzymes play a major role in scavenging reactive oxygen species that arise during somatic embryogenesis (SE). Activity levels of antioxidant enzymes such as peroxidase (POX), catalase (CAT), and superoxide dismutase (SOD) were investigated during developmental stages of SE of Musa acuminata Colla (AAA group) ‘Grand Naine’ and Musa spp. (AAB group) ‘Rasthali’. Using native polyacrylamide gel electrophoresis, five POX isoforms were detected in ‘Rasthali’, and four POX isoforms were observed in ‘Grand Naine’. The POX1 isoform was highly expressed in non-embryogenic callus (NEC) of both cultivars, and the POX2 isoform was present in somatic embryos (se) at 0, 30, 45, and 60 d of development of ‘Rasthali’, but not ‘Grand Naine’. Among two CAT isoforms, the CAT2 isoform showed higher expression in embryonic stages than in vegetative stages in explants and NEC of both cultivars. A total of five SOD isoforms detected in different developmental stages of both cultivars, SOD4 and SOD5, were detected only during the developmental stages of se. The enzyme assay showed that the highest CAT and SOD activity was in embryogenic callus (EC) of both cultivars, whereas activity of POX was reduced in EC. During somatic embryo development, the activity of SOD, POX, and CAT was found to be higher in the early globular stage than in the later coleoptilar stage embryos. During germination, the activity of SOD and POX was found to be higher in germinating se of ‘Grand Naine’. The specifically detected isoforms can be used as biochemical markers.ICAR-NRC

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Not AvailableAvailability of transcriptome datasets for use in accelerated molecular-based breeding in Musa species is limited. Illumina Hiseq technology was employed to determine differential gene expression between the contrasting cultivars for three different stresses (Eumusae leaf spot –Mycosphaerella eumusae, root lesion nematode – Pratylenchus coffeae and moisture deficit stress) under challenged and unchallenged conditions. An average of 34.72 million of reads was assembled into *47629 contigs, and *5,466 simple sequence repeats (SSR) from each library were identified. GO annotation and KEGG pathway analysis were carried for all the transcripts and the SSR, SNPs were also detected. Based on this information, a MusatransSSRDB has been developed. Currently, the database consists of 32,800 SSRs with the unique information like putative function of the SSR-containing genes and their metabolic pathway and expression profiling under various stress conditions. This database provides information on in silico polymorphic SSRs (2830 SSRs) between the contrasting cultivars for each stress and within stress. Information on in silico polymorphic SSRs specific to differentially expressed genes under challenged condition for each stress can also be accessed. This database facilitates the retrieval of results by navigating the tabs for cultivars, stress and polymorphism. This database was developed using HTML, Java and PHP; datasets are stored in MySQL database and accessible in the public domain (http://bioinfnrcb.byethost7.com/nrcbbio/). This unique information facilitates the banana breeder to select the SSR primers based on specific objectives. MusatransSSRDBalong with other genomics databases will facilitate the genetic dissection and breeding for complex traits in banana. Thus,this database is a step forward in economizing cost, time, manpower and other resources.ICAR, NPTC–Functional Genomics

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Not AvailableGermination of somatic embryos (se) is an important event in a whole somatic embryogenesis (SE) process. Though the well matured somatic embryos of banana were placed on the germination medium with complete contact only 50% of the somatic embryos were germinated into plantlets having either shoots, roots or sometimes both. Hence in order to increase the germination efficacy it is essential to understand the molecular basis underlying the process of germination. In the present study, we have analyzed differential proteome expression between germinating somatic embryo (Gse) and non germinating somatic embryos (NGse) of cv. Grand Naine by using two dimensional gel electrophoresis followed by MALDI-TOF MS/MS analysis. In total 16 protein spots were differentially expressed between Gse and NGse in which 15 were successfully identified. In that, 14 spots were upregulated and 1 spot was downregulated in Gse than NGse. High accumulation of SAUR auxin responsive proteins and cold stress proteins like 3-ketoacyl-CoA synthase and Oleosin were highly expressed in Gse than NGse. Hence SAUR triggering factors like NAA, and GA3 were supplemented in the germination medium in three combinations along with cold treatments of 4°C at 0 hr, 12 hr and 24 hr. High germination efficiency of 93% was observed in 24 hr treated embryos in medium supplemented with 1mg/L NAA and 0.5 mg/L GA3.Not Availabl

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Not AvailableSomatic embryogenesis is an important tool for crop improvement through transgenic approach and even for gene editing. It has been hypothesized regularly for large-scale propagation of banana which necessitates basic data on genetic fidelity and field performance of the plants towards ensure the commercial feasibility of the technique. Plantlets regenerated from embryogenic cell suspension (ECS) cultures established using immature male flower buds were examined for genetic fidelity using Inter Simple Sequence Repeats (ISSR) markers. Results showed that the primers UBC 808, UBC 811 and UBC 841 each generated one polymorphic band with an overall variation in banding pattern of 3.34 and 2.09% in cvs. Grand Naine and Rasthali respectively. Field evaluation of the ECS derived plants showed that there were no negative effects on the vegetative and yield parameters. Remarkably no phenotypic off-types were observed in this field trial. The level of genetic variation observed in this study is not an obstacle for further uptake of this novel propagation technique.Not Availabl

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Not AvailableLong non-coding RNAs (lncRNAs) are important regulators of gene expression in plant response to biotic and abiotic stresses. The participation of lncRNAs in plant disease resistance in bananas is largely unknown. Therefore, we attempted to identify novel lncRNAs responsive to Mycosphaerella eumusae, a causative agent of Eumusae leaf spot disease and root lesion nematode, Pratylenchus coffeae and their differential expression patterns during stress and normal conditions in respective resistant and sensitive banana genotypes. Illumina paired-end transcriptome sequencing of control and infected samples of resistant and sensitive banana cultivars was performed and the sequence reads assembled into 172434, 201256 transcriptional units (TUs) for M. eumusae and P. coffeae, respectively. The genome-wide analysis for ELSD-responsive lncRNAs led to identification of 5142 novel lncRNAs including 3031-intergenic, 1672-intragenic, and 440 antisense lncRNAs classes collectively from ELSD-resistant and sensitive cultivars. Similarly, 5615 lncRNAs comprised of 3283 intergenic, 1878-intragenic, and 454 antisense classes were identified from P. coffeae-infected resistant and sensitive banana cultivars. Most of the lncRNAs were stress specific, evenly distributed among banana chromosomes and the average length is ranging from 620 to 684 nucleotides. In addition, 1250 and 1284 lncRNAs were differentially expressed to M. eumusae and P. coffeae infections, respectively. The LncRNA–mRNA interaction-based functional role showed lncRNA-mediated downregulation of horcolin, an antifungal protein is likely responsible for ELSD sensitivity in banana cultivars. Furthermore, we identified 100 of these lncRNAs also play a role in drought stress response of banana indicating a possible crosstalk between biotic and abiotic stresses.NPTC-Functional Genomics component of the Indian Council of Agricultural Research (ICAR), India

Drought Stress-Modulated Alternative Splicing Landscapes in Drought-tolerant and -Sensitive Banana Cultivars

Not Availablevariants have major impact in plant response to drought stress. Alternative splicing a major post-transcriptional modification and its differential sensitivity to drought stress is of paramount importance to resolve complex molecular response of drought stress and to develop drought-resilient crops. In the present study, we analyzed the alternative splicing pattern of drought-tolerant (DT) and -sensitive banana (DS) cultivars under drought conditions and found that the number of spliced transcripts in DS (AAA genome) has increased to about 4.32 folds, while 0.19 fold among all the splicing events were reduced in DT (ABB genome). Categorization of drought -modulated alternative splicing (AS) events revealed that intron retention is the most abundant (42.5%) process, followed by alternative splice acceptor (22.6%), alternative splice donor (12.2%), and exon skipping (4.86%) in DS. Only 40-44% of intron retained transcripts have the protein coding capacity, indicating that their occurrence would participate in drought stress response as modified yet functional proteins. We observed that retained introns were slightly higher in GC content than constitutively spliced introns. The results reveal that the genotype dependent AS pattern may play an important role in drought tolerance in banana. This study will help in deciphering the molecular basis underlying phenotypic differences among tolerant and sensitive banana cultivars.Not Availabl

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Not AvailableIn banana, drought responsive gene expression profiles of drought-tolerant and sensitive genotypes remain largely unexplored. In this research, the transcriptome of drought-tolerant banana cultivar (Saba, ABB genome) and sensitive cultivar (Grand Naine, AAA genome) was monitored using mRNA-Seq under control and drought stress condition. A total of 162.36 million reads from tolerant and 126.58 million reads from sensitive libraries were produced and mapped onto the Musa acuminata genome sequence and assembled into 23,096 and 23,079 unigenes. Differential gene expression between two conditions (control and drought) showed that at least 2268 and 2963 statistically significant, functionally known, non-redundant differentially expressed genes (DEGs) from tolerant and sensitive libraries. Drought has up-regulated 991 and 1378 DEGs and down-regulated 1104 and 1585 DEGs respectively in tolerant and sensitive libraries. Among DEGs, 15.9% are coding for transcription factors (TFs) comprising 46 families and 9.5% of DEGs are constituted by protein kinases from 82 families. Most enriched DEGs are mainly involved in protein modifications, lipid metabolism, alkaloid biosynthesis, carbohydrate degradation, glycan metabolism, and biosynthesis of amino acid, cofactor, nucleotide-sugar, hormone, terpenoids and other secondary metabolites. Several, specific genotype-dependent gene expression pattern was observed for drought stress in both cultivars. A subset of 9 DEGs was confirmed using quantitative reverse transcription-PCR. These results will provide necessary information for developing drought-resilient banana plants.Not Availabl

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Not AvailableEndogenous hormone secretion proteins along with stress and defense proteins play predominant role in banana embryogenesis. This study reveals the underlying molecular mechanism during transition from vegetative to embryogenic state.ICAR, ICAR-NRC

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Not AvailableEvaluation of 100 Indian Musa accessions (IMA) for nine elements in their fresh fruit pulp (FFP) revealed genetic variability of 4.7-fold for K & Mg to 111.1-fold for Ca but, only with either highly or moderately positively skewed distribution. The descending order of mineral concentrations (MC) was K > Ca > Na > Mg > Fe > Mn > B > P > Zn. 100 g FFP contributes fairly about 5 (Fe) to 10% (Mn, Ca & Mg) of daily mineral requirement of Indians. Calcium (97%) and Fe (96%) showed the highest heritability while Zn exhibited lowest (85%). Significantly positive correlation was observed for all minerals. Magnesium had maximum direct effect on Fe content followed by Mn, Zn and Na in path analysis. Both principal component analysis and cluster analysis failed to group the IMA according to their ploidy/genome/subgroups. Twenty commercial cultivars were placed in top 10 positions based on their MC. Besides Ca and Mg, IMA were richer for all micronutrients than the world’s Musa gene-pool.Not Availabl

Inter retrotransposon based genetic diversity and phylogenetic analysis among the Musa germplasm accessions

Not AvailableIn the present investigation, the insertional polymorphisms of retro-elements were studied in the Musa germplasm available at ICAR-NRCB field gene bank using IRAP markers. The maximum number of polymorphic bands were produced by the primer pair Nikita and LTR 6150 (48) followed by LTR 6149 and 3′LTR (47) and minimum of 35 bands were produced by the primer pair Sukkula and LTR 6150. The bands produced were scored as 0 (absent) and 1 (present) and the resultant binary data was subjected to diversity analysis. The dendrogram consisted of two major clusters with members of Eumusa and Rhodochlamys in one indicating their genetic closeness and members of the genus Ensete in another cluster. Results of principal coordinate analysis were congruent to those obtained in hierarchial cluster analysis. The molecular markers used in this study could reveal intra and inter-group diversity among the Musa germplasm accessions with similarity co-efficient ranging from 0.41 to 0.99. IRAP marker system has performed excellently clustering the accessions based on both genomic and subgroup levels. The entire germplasm was found to be robust with no duplications indicating the diverse group of accessions available at ICAR-NRCB field gene bank. It has also exhibited high polymorphism and hence could be effectively used to detect the genetic relatedness among diverse genome of Musa.Not AvailableICAR, GoI, Indi