Brassica biodiversity conservation: prevailing constraints and future avenues for sustainable distribution of plant genetic resources

The past decade has seen an observable loss of plant biodiversity which can be attributed to changing climate conditions, destroying ecosystems to create farmlands and continuous selective breeding for limited traits. This loss of biodiversity poses a significant bottleneck to plant biologists across the globe working on sustainable solutions to address the current barriers of agricultural productivity. Plant genetic resources centers or genebanks that conserve plant germplasm can majorly contribute towards addressing this problem. Second only to soybean, Brassica remains the largest oil-seed crop and is cultivated across 124 countries, and FAO estimates for a combined gross production values of broccoli, cabbages, cauliflower, mustard and rape seeds stands at a staggering 67.5 billion US dollars during the year 2020. With such a global status, wide variety of uses and more recently, growing importance in the health food sector, the conservation of diverse genetic resources of Brassica appeals for higher priority. Here we review the current status of Brassica conservation across plant genebanks. At present, at least 81,752 accessions of Brassica are recorded to be conserved in 148 holding institutes spread across only 81 countries. Several aspects that need to be addressed to improve proper conservation of the Brassica diversity was well as dissemination of germplasm are discussed. Primarily, the number of accessions conserved across countries and the diversity of Brassica taxa most countries has been highly limited which may lead to biodiversity loss in the longer run. Moreover, several practical challenges in Brassica germplasm conservation especially with respect to taxonomic authorities have been discussed. The current review identifies and highlights areas for progress in Brassica conservation, which include but are not limited to, distribution of conserved Brassica biodiversity, challenges faced by conservation biologists, conservation methods, technical hurdles and future avenues for research in diverse Brassica species

Differential Metabolic Profiles during the Developmental Stages of Plant-Parasitic Nematode Meloidogyne incognita

Meloidogyne incognita is a common root-knot nematode with a wide range of plant hosts. We aimed to study the metabolites produced at each stage of the nematode life cycle to understand its development. Metabolites of Meloidogyne incognita were extracted at egg, J2, J3, J4, and female stages and 110 metabolites with available standards were quantified using CE-TOF/MS. Analyses indicated abundance of stage-specific metabolites with the exception of J3 and J4 stages which shared similar metabolic profiles. The egg stage showed increased abundance in glycolysis and energy metabolism related metabolites while the J2 metabolites are associated with tissue formation, motility, and neurotransmission. The J3 and J4 stages indicated amino acid metabolism and urea cycle- related metabolites. The female stage was characterized with polyamine synthesis, antioxidant activity, and synthesis of reproduction related metabolites. Such metabolic profiling helps us understand the dynamic physiological changes related to each developmental stage of the root-knot nematode life cycle

Stage-Wise Identification and Analysis of miRNA from Root-Knot Nematode Meloidogyne incognita

In this study, we investigated global changes in miRNAs of Meloidogyne incognita throughout its life cycle. Small RNA sequencing resulted in approximately 62, 38, 38, 35, and 39 Mb reads in the egg, J2, J3, J4, and female stages, respectively. Overall, we identified 2724 known and 383 novel miRNAs (read count > 10) from all stages, of which 169 known and 13 novel miRNA were common to all the five stages. Among the stage-specific miRNAs, miR-286 was highly expressed in eggs, miR-2401 in J2, miR-8 and miR-187 in J3, miR-6736 in J4, and miR-17 in the female stages. These miRNAs are reported to be involved in embryo and neural development, muscular function, and control of apoptosis. Cluster analysis indicated the presence of 91 miRNA clusters, of which 36 clusters were novel and identified in this study. Comparison of miRNA families with other nematodes showed 17 families to be commonly absent in animal parasitic nematodes and M. incognita. Validation of 43 predicted common and stage-specific miRNA by quantitative PCR (qPCR) indicated their expression in the nematode. Stage-wise exploration of M. incognita miRNAs has not been carried out before and this work presents information on common and stage-specific miRNAs of the root-knot nematode

Glucosinolate Diversity Analysis in Choy Sum (<i>Brassica rapa</i> subsp. <i>chinensis</i> var. <i>parachinensis</i>) Germplasms for Functional Food Breeding

The aim of this study was to analyze glucosinolates (GSLs) in germplasm that are currently conserved at the RDA-Genebank. The analysis focused on the glucosinolate diversity among the analyzed germplasms, with the goal of identifying those that would be most useful for future breeding efforts to produce nutritionally rich Choy sum plants. In total, 23 accessions of Choy sums that possessed ample background passport information were selected. On analyzing the glucosinolate content for 17 different glucosinolates, we observed aliphatic GSLs to be the most common (89.45%) and aromatic GSLs to be the least common (6.94%) of the total glucosinolates detected. Among the highly represented aliphatic GSLs, gluconapin and glucobrassicanapin were found to contribute the most (>20%), and sinalbin, glucoraphanin, glucoraphasatin, and glucoiberin were detected the least (less than 0.05%). We identified one of the accessions, IT228140, to synthesize high quantities of glucobrassicanapin and progoitrin, which have been reported to contain several therapeutic applications. These conserved germplasms are potential bioresources for breeders, and the availability of information, including therapeutically important glucosinolate content, can help produce plant varieties that can naturally impact public health

High-Throughput Phenotypic Characterization and Diversity Analysis of Soybean Roots (<i>Glycine max</i> L.)

Soybean (Glycine max L.) is a crop native to Northeast Asia, including China, Korea, and Japan, but currently cultivated all over the world. The National Agrobiodiversity Center in Korea at the Rural Development Administration (RDA) conserves approximately 26,000 accessions and conducts characterizations of its accessions, to accumulate new information. Roots are essential organs of a plant, providing mechanical support, as well as aiding water and nutrient acquisition. Currently, not much information is available in international gene banks regarding root characterization. We studied the root phenotype of 374 soybean accessions, using a high-throughput method. Eight root morphological traits (RMT) were studied and we observed that the surface area (SA), number of forks (NF), and number of tips (NT) had a positive correlation with total length (LENGTH), and that link average length (LAL) and other traits all had a negative correlation. Additionally, the correlation between seed traits (height, width, and 100-seed weight) and root traits was confirmed for the first time in this experiment. The germplasms were divided into three clusters by k-means clustering, and orthogonal projections to latent structures discriminant analysis (OPLS-DA) was used to compare clusters. The most distinctive characteristics between clusters were total lateral average length (LAD) and total lateral average length (DIAM). Cluster 3 had the highest LENGTH, SA, NF, and NF, whereas cluster 1 had the smallest LENGTH, SA, and NF. We selected the top 10 accessions for each RMT, and IT208321, IT216313, and IT216137 were nominated as the best germplasms. These accessions can be recommended to breeders as materials for breeding programs. This is a preliminary report on the characterization of the root phenotype at an international gene bank and will open up the possibility of improving the available information on accessions in gene banks worldwide

Exogenous expression of ACC deaminase gene in psychrotolerant bacteria alleviates chilling stress and promotes plant growth in millets under chilling conditions

463-468Endogeneous ethylene evolved during cold stress is a major limiting factor for plant growth which can be controlled by bacterial enzyme 1-aminocyclopropane-1-carboxylate (ACC) deaminase (ACCD), by breaking down ACC, the precursor of ethylene. In the present study, we introduced ACCD expressing plasmid in ACCD negative psychrotolerant bacteria to study its effect on growth of finger and foxtail millet seedlings. ACCD negative Sphingomonas faeni ISO were selected and transformed with plasmid pRKACC containing the acdS gene. Inoculation of the millet seeds and studying physiological parameters when a cold stress of 4 and 10ºC was imposed showed that inoculation with ACCD expressing strains improved root and shoot length, biomass content of foxtail and finger millets seeds. Further, we also observed increased antioxidant activity in the plants by high levels of SOD, CAT, GPX, POD, APX and GR enzyme activity, and decreased proline content on inoculation with ACCD positive mutants. The enzyme ACC deaminase is thus be proved to be a potential strategy to alleviate cold stress in foxtail and finger millet by regulating endogenous ethylene evolved during stress conditions

Isolation and characterization of nanocellulose from selected hardwoods, viz., Eucalyptus tereticornis Sm. and Casuarina equisetifolia L., by steam explosion method

Abstract Extraction of nanocellulose is challenging, especially from hardwoods due to its complex chemical structure as well as structural hierarchy. In this study, nanocellulose was isolated from wood pulp of two hardwood species, namely Eucalyptus tereticornis Sm. and Casuarina equisetifolia L. by steam explosion process. Pure cellulose wood pulp was obtained through Kraft pulping process followed by alkaline and bleaching pre-treatments. Isolated nanocellulose was characterized by Scanning Electron Microscopy (SEM), Transmission Electron Microscopy (TEM), Atomic Force Microscopy (AFM), Fourier Transformed Infrared (FTIR) Spectra, Thermogravimetric Analysis (TGA), and X-ray diffraction (XRD) studies. Nanocellulose obtained from both species showed non-significant difference with average diameter of 27.801 nm for eucalyptus and 28.690 nm for casuarina, which was confirmed from TEM and AFM images. FTIR spectra of nanocellulose showed prominent peaks corresponding to cellulose and absence of peaks corresponding to lignin. The elemental purity of nanocellulose was confirmed with EDAX detector. XRD analysis showed the enrichment of crystalline cellulose in nanocellulose, and also confirmed the significant conversion of cellulose I to cellulose II. During TG analysis the untreated fibres started to degrade earlier than the nanocellulose which indicated the higher thermal stability of nanocellulose. Highly entangled network like structure along with high aspect ratio make the nanofibres a versatile material for reinforcing the composites. This successful method can be replicated for industrial level production of cellulose nanofibres

Optimization of Extraction Process and Kinetics of <i>Sterculia foetida</i> Seed Oil and Its Process Augmentation for Biodiesel Production

This article reports optimization and kinetic studies on extraction of <i>Sterculia foetida</i> seed oil and process optimization for biodiesel production from the same. The oil extraction follows first-order kinetics, and the yield was found to reach a maximum of 55.58 wt % for a 1:12 seed-to-hexane weight ratio. The activation energy and activation thermodynamic parameters at 338 K were determined as <i>E</i>_a = 69.441 kJ mol^–1, Δ<i>H</i>^‡ = 66.63 kJ mol^–1, Δ<i>S</i>^‡ = −238.07 J mol^–1 K^–1, and Δ<i>G</i>^‡ = 147.09 kJ mol^–1. Complete physicochemical properties of the oil were analyzed using standard methods. The low acid value of 0.42 mg of KOH g^–1 for fresh oil enables alkali catalytic transesterification. Different biodiesel production parameters including methanol-to-oil molar ratio, catalyst concentration, and reaction temperature were examined. An optimum yield of 95.4 wt % with a conversion of 98.91% was achieved at values of 6:1, 0.9 wt %, and 338 K, respectively. The fuel properties of the produced biodiesel were compared with the ASTM D6751 biodiesel standard