Kishan Saha

Bananas and plantains, belonging to the Musa genus, are important food crops that sustain the livelihoods of countless smallholder farmers globally. However, their production is hindered by various challenges, including abiotic and biotic stresses, climate change, and poor access to clean planting materials, which negatively impact their yields. Addressing these constraints is essential for improving production and ensuring food security. This study investigated the influence of triploid genome background and exogenous growth regulators on the regeneration of Musa cultivars [Gros Michel (AAA genome), Obino l’Ewai and Silk (AAB genome), and Poteau Naine (ABB genome)]. Shoot tip explants of the AAA, AAB, and ABB triploid genomes were cultured in Murashige and Skoog (MS) media supplemented with varying 6-benzylaminopurine (BAP) and indole-3-butyric acid (IBA), indole-3-acetic acid (IAA), or naphthaleneacetic acid (NAA) hormones. Shoot induction was successfully achieved within 21.50 ± 2.00 days, with AAA exhibiting the highest shoot induction frequencies ranging from 30.00 ± 1.57% to 100% and shoot numbers per explant ranging from 3.00 ± 0.50 to 8.80 ± 0.80, followed by the ABB genome ranging from 20.00 ± 3.45% to 100% and from 2.00 ± 0.55 to 5.60 ± 0.50 shoots, and the AAB genome ranging from 17.50 ± 5.01% to 100% and from 2.00 ± 0.04 to 6.60 ± 0.25 shoots, respectively, in media amended with 1.2 to 6.0 mg.L⁻¹ BAP and 0.1 mg.L⁻¹ IAA. The highest rooting rate of 100% was recorded in all three genomes in media containing 1.4 mg.L⁻¹ IBA and 0.5 mg.L⁻¹ IAA, with the AAA genome producing the maximum number of 14.8 roots per explant. The results indicate the positive influence of the AAA genome background on in vitro regeneration and its potential utilization for genomic editing transformation protocols

Small non-coding microRNAs (miRNAs) play crucial roles in the degradation of the messenger RNAs (mRNAs) that are involved in various biological processes post-transcriptionally and translationally. Many plants, especially Musa spp. (plantains and bananas), which are important perennial herbs of the family Musaceae, experience significant yield loss due to abiotic stressors, yet only a few miRNAs involved in this response have been identified. This study employed in silico analyses of transcriptome shotgun assembly (TSA) and expressed sequence tag (EST) sequences to identify Musa miRNAs and their target genes. Leaf and root tissues from three Musa genomic groups (AAA, AAB, and ABB) under drought stress were analyzed using quantitative real-time PCR (qRT-PCR) to validate the expression of miRNAs. A total of 17 potential conserved miRNAs from 11 families were identified, with the minimal folding free energies (-kcal/mol) of precursors ranging from −136.00 to −55.70, as observed through RNA folding analysis. Six miRNAs (miR530-5p, miR528-5p, miR482a, miR397a, miR160h, and miR399a) showed distinct tissue-specific expression patterns in the roots and leaves across the three groups. A total of 59 target regulatory transcription factors and enzymes involved in stress response, growth, and metabolism were predicted. Of these, 11 targets were validated for miR530-5p, miR528-5p, miR482a, and miR397a, using qRT-PCR. These four stress-responsive miRNAs exhibited an inverse expression relationship with their target genes across two different tissues in Musa groups. This research provides insights into miRNA-mediated drought stress responsiveness in Musa spp., potentially benefiting future studies on gene regulation under drought stress.

The high quantity of polyphenols and polysaccharides present in the tissues of Musa spp. often leads to the degradation of nucleic acids, which is why all previously established methods resulted in lesser yield and poor quality of RNA. In this study, we present a modified SDS-based RNA extraction method to improve the quality and yield of RNA from different tissues of Musa spp. for downstream applications. The modification of RNA extraction buffer, SDS, heat incubation, and use of LiCl resulted in high-intensity RNA bands and increased RNA yield. The clear ribosomal RNA bands ensured the high quality of intact RNA without genomic DNA contamination, along with A260/A280 and A260/A230 ratios ranging from 1.83 to 2.25, which indicated the high quality of RNA across different banana varieties and tissue types. This method was found to be very effective when RNA was extracted from drought-stressed plants yielding 2.92 to 6.30 µg/100 mg fresh weight with high RNA integrity and quality (RNA IQ) 7.8–9.9 from the different groups of Musa tissues. Additionally, the RNA was successfully applied in PCR and quantitative real-time PCR (qRT-PCR), confirming downstream application in gene expression analysis. This method is a reliable and efficient technique for RNA extraction methods like Trizol, NucleoSpin, RNeasy, and CTAB procedures reported so far.

Phased short-interfering RNAs (phasiRNAs) fine-tune various stages of growth, development, and stress responses in plants. Potato (Solanum tuberosum) tuberization is a complex process, wherein a belowground modified stem (stolon) passes through developmental stages like swollen stolon and mini-tuber before it matures to a potato. Previously, we identified several phasiRNA-producing loci (PHAS) from stolon-to-tuber transition stages. However, whether phasiRNAs mediate tuber development remains unknown. Here, we show that a gene encoding NB-ARC DOMAIN-CONTAINING DISEASE RESISTANCE PROTEIN (StRGA4; a PHAS locus) is targeted by Stu-microRNA482c to generate phasiRNAs. Interestingly, we observed that one of the phasiRNAs, referred as short-interfering RNA D29 (-) i.e. siRD29 (-), targets the gibberellin (GA) biosynthesis gene GIBBERELLIN 3-OXIDASE 3 (StGA3ox3). Since regulation of bioactive GA levels in stolons controls tuber development, we hypothesized that a gene regulatory module, Stu-miR482c-StRGA4-siRD29(-)-StGA3ox3, could govern tuber development. Through transient expression assays and small RNA-sequencing, generation of siRD29(-) and its phase was confirmed in planta. Notably, the expression of StGA3ox3 was higher in swollen stolon compared to stolon, whereas siRD29(-) showed a negative association with StGA3ox3 expression. Antisense (AS) lines of StGA3ox3 produced more tubers compared to wild-type.. As expected, StRGA4 overexpression (OE) lines had high levels of siRD29(-) and mimicked the phenotypes of StGA3ox3-AS lines, indicating the functionality of this module in potato. In vitro tuberization assays (with or without GA inhibitor) using StGA3ox3 antisense lines and overexpression lines of StGA3ox3 or StRGA4 revealed that StGA3ox3 controls the tuber-stalk development. Taken together, our findings suggest that a phasiRNA, siRD29(-), mediates the regulation of StGA3ox3 during stolon-to-tuber transitions in potato.

Plants exhibit diverse developmental plasticity and modulate growth responses under various environmental conditions. Potato (Solanum tuberosum), a modified stem and an important food crop, serves as a substantial portion of the world’s subsistence food supply. In the past two decades, crucial molecular signals have been identified that govern the tuberization (potato development) mechanism. Interestingly, microRNA156 overexpression in potato provided the first evidence for induction of profuse aerial stolons and tubers from axillary meristems under short-day (SD) photoperiod. A similar phenotype was noticed for overexpression of epigenetic modifiers—MUTICOPY SUPRESSOR OF IRA1 (StMSI1) or ENAHNCER OF ZESTE 2 (StE[z]2), and knockdown of B-CELL-SPECIFIC MOLONEY MURINE LEUKEMIA VIRUS INTEGRATION SITE 1 (StBMI1). This striking phenotype represents a classic example of modulation of plant architecture and developmental plasticity. Differentiation of a stolon to a tuber or a shoot under in vitro or in vivo conditions symbolizes another example of organ-level plasticity and dual fate acquisition in potato. Stolon-to-tuber transition is governed by SD photoperiod, mobile RNAs/proteins, phytohormones, a plethora of small RNAs and their targets. Recent studies show that polycomb group proteins control microRNA156, phytohormone metabolism/transport/signaling and key tuberization genes through histone modifications to govern tuber development. Our comparative analysis of differentially expressed genes between the overexpression lines of StMSI1, StBEL5 (BEL1-LIKE transcription factor [TF]), and POTATO HOMEOBOX 15 TF revealed more than 1,000 common genes, indicative of a mutual gene regulatory network potentially involved in the formation of aerial and belowground tubers. In this review, in addition to key tuberization factors, we highlight the role of photoperiod and epigenetic mechanism that regulates the development of aerial and belowground tubers in potato.

Tripura being a state in North East India belongs to Indo-Burma bio-diversity hotspot and is considered as a centre of origin of many species of Zingiberaceae. Alpinia calcarata, Alpinia malaccensis, Alpinia nigra, Amomum aromaticum, Amomum koenigii, Amomum maximum, Curcuma amada, Curcuma caesia, Curcuma longa, Curcuma picta, Hedychium coccineum, Hedychium coronarium and Hedychium thyrsiforme are found in wild state in different geographical locations of Tripura. Their karyotypes were analyzed both at interspecific and intraspecific levels. The somatic chromosome number of Alpinia spp. and Amomum spp. was found to be 2n = 4X = 48. The Curcuma spp. represented by C. amada had 2n = 42 chromosomes and three other species viz., C. caesia, C. longa and C. picta had 2n = 63 chromosomes having X=21, indicating that polyploidy is a common feature in this genus. The somatic chromosome number of Hedychium spp. was found to be 2n = 34 chromosomes having a basic no. X = 17. Chromosomal data based clustering pattern and their sub-grouping at intra-specific level validates the taxonomic status of these species. Gower’s similarity matrix is an indicator of cryptic changes leading genus specific karyotype conservatism.

Polymerase Chain reaction (PCR) based molecular characterization has been undertaken for assessing the genetic diversity in five populations of Solanum torvum using SSR and RAPD markers. In this study, 8 SSRs produced 151 fragments of which 131 bands were polymorphic (86.38%). The primers, At5 amplified the highest number of polymorphic loci (27) and the highest PIC was recorded in CBT08 (0.54). In comparison, RAPD assay produced 70 bands with 79.16% polymorphism. The PIC value was highest in OPC14 (0.41). UPGMA clustering for SSR and RAPD markers grouped all the populations into two clusters. Our findings on SSR profile suggests that though different populations of S. torvum are inherited from a common ancestor eventually the population (STP5) with greater genetic diversity is stabilized in the high altitude of Sub - Himalayan region of Tripura in the due course of evolution.

This review has been mainly focussed on distribution, cytology, genetic diversity and molecular phylogeny of selected species of Zingiberaceae belonging to four genera of high economic importance viz., Alpinia, Amomum, Curcuma and Hedychium. Zingiberaceae comprising ca.50 genera and more than 1600 species is mostly distributed in tropical and subtropical areas. Cytological and genetic diversity analyses so far carried out obviously, indicated the genetic relatedness or variation of the species both at intergeneric and intrageneric levels. The DNA barcode and molecular phylogeny also point towards the degree of phylogenetic relationships between the species and also supplement the details of preferred barcode candidates for precise identification of Zingiberean species.

The present investigation has been focussed on the inter-chromosomal and intra-chromosomal asymmetry data of Allium sativum and Scadoxus multiflorus of Amaryllidaceae using quali-quantitative and quantitative methods. The quail-quantitative analysis is strictly ratio based and it revealed that the karyotype of A. sativum and S. multiflorus belonged to category 1A and 3C respectively indicating only the degree of asymmetry. The coefficient of variation of chromosome length (CVCL) and the mean centromeric asymmetry (MCA) were the estimators of quantitative analysis and the data showed that the inter- and intra-chromosomal asymmetry were comparatively high in S. multiflorus than that of A. sativum. Further, the genetic distance between these two species is clearly visible in the bi-dimensional scattered plot from the derived MCA and CVCL data.