Nutraceutical and flavor profiles in underutilized desert legumes of India: gene editing strategies towards sustainable food development.

Climate change has posed a challenge for food security all over the world in the form of fluctuating crop yields and novel disease outbreaks in plants. Human society's overdependence on a few food crops does not seem a wise precedence. There are numerous underutilized/orphan/neglected legumes growing in the Indian desert regions that can come to the rescue and act as balanced and sustainable sources of nutrients and health-benefitting nutraceuticals. However, challenges such as low plant yield, unidentified metabolic pathways and off-flavor in the food products derived from them prevent the realization of their full potential. Conventional breeding techniques are too slow to achieve the desired modifications and cater to the sharply rising demand for functional foods. The novel gene editing tools like CRISPR-Cas provide more precise tool to manipulate the target genes with or without introduction of foreign DNA and therefore, have better chances to be accepted by governments and societies. The current article reports some of the relevant 'gene editing' success stories with respect to nutraceutical and flavor profiles in the popular legumes. It highlights gaps and future potential, along with areas requiring caution, in underutilized edible legumes of the Indian (semi) arid regions like Prosopis cineraria, Acacia senegal and Cyamopsis tetragonoloba.

Adventitious root formation in crops-Potato as an example.

The root system of potato is made up of adventitious roots (AR) that form at the base of a sprout once it emerges from the mother tuber. By definition, AR originate from dormant preformed meristems, or from cells neighboring vascular tissues in stems or leaves. This may occur as part of the developmental program of the plant (e.g., potato), or when replacing the embryonic primary roots in response to stress conditions, such as flooding, nutrient deprivation, or wounding. AR formation is studied mainly in cereals and model plants, and less is known about its developmental program in root and tuber crops. In this review, we summarize the recent data on AR development in potato and relate this knowledge to what is known from model plants. For example, AR formation following stem cutting in potato follows a pattern of initiation, expression, and emergence phases that are known for other plants and involves auxin, the master regulator of AR induction and development. Molecular regulation of AR formation and the effect of environmental stresses are discussed. Understanding the origin and nature of AR systems in important crops will contribute to increased production and improve global food security.

Cloning and characterization of the Salicornia brachiata Na(+)/H(+) antiporter gene SbNHX1 and its expression by abiotic stress.

Salinity causes multifarious adverse effects to plants. Plants response to salt stress involves numerous processes that function in coordination to alleviate both cellular hyperosmolarity and ion disequilibrium. A Na(+)/H(+) antiporter NHX1 gene has been isolated from a halophytic plant Salicornia brachiata in this study. Predicted amino acid sequence similarity, protein topology and the presence of functional domains conserved in SbNHX1 classify it as a plant vacuolar NHX gene. The SbNHX1 cDNA has an open reading frame of 1,683 bp, encoding a polypeptide of 560 amino acid residues with an estimated molecular mass 62.44 kDa. The SbNHX1 shows high amino acid similarity with other halophytic NHX gene and belongs to Class-I type NHXs. TMpred suggests that SbNHX1 contains 11 strong transmembrane (TM). Real time PCR analysis revealed that SbNHX1 transcript expresses maximum at 0.5 M. Transcript increases gradually by increasing the treatment duration at 0.5 M NaCl, however, maximum expression was observed at 48 h. The overexpression of SbNHX1 gene in tobacco plant showed NaCl tolerance. This study shows that SbNHX1 is a potential gene for salt tolerance, and can be used in future for developing salt tolerant crops.

Pomegranate Fruit Growth and Skin Characteristics in Hot and Dry Climate.

Pomegranate (Punica granatum L.) fruit is well known for its health-beneficial metabolites. The pomegranate peel consists of an inner thick spongy white tissue, and an outer smooth skin layer that accumulates anthocyanins in red cultivars when ripe. The skin is made up of epidermis cells covered by a cuticle, the latter being the first target of cracking and russeting. The present study focuses on the effect of Israel's hot and dry climate on pomegranate growth, to elucidate the derived effects on fruit skin characteristics and its putative resistance to the building pressure from fruit expansion. Experiments were conducted for four years, in four orchards located in different regions of the country, each with a different typical microclimate. Fruit-growth parameters were followed using remote-sensing tools, microscopic study, and mineral analysis of the skin, followed by determination of the peel's elastic modulus. Fruit expanded in two phases: a short rapid phase followed by a gradual phase with a sigmoidal growth-rate pattern. Extreme hot and dry climate during the period of maximal growth rate was associated with restricted growth and a high proportion of small-size fruit. Anatomical study indicated that the skin of mature pomegranate fruit is made up of epidermal cells that are relatively flat and spaced apart, and is expected to be less durable against internal pressure. In contrast, skin of early immature fruit has two layers of dense and rounded epidermis, and is expected to be more resistant to cracking. Tensile strength studies confirmed this trend-skin of mature fruit had a lower elastic modulus than young fruit. However, restrained growth due to extreme environmental cues may result in better resistance of the mature pomegranate fruit to cracking, and in better skin quality and appearance, albeit small fruits. On the other hand, temperate climate at the beginning of the growth period, which allows high growth rate and high daily shrinkage, leads to pomegranate skin disorders.

Identification of candidate genes mediating apple fruit-cracking resistance following the application of gibberellic acids 4 + 7 and the cytokinin 6-benzyladenine.

Calyx-end cracking in 'Pink Lady' apple is treated by a solution of gibberellic acids 4 and 7 (GA4+7) and the cytokinin 6-benzyladenine (BA). Although the GA4+7 and BA mixture is applied early in apple fruit development, it mitigates cracking that becomes evident in the mature fruit, implying a long-term treatment effect. The reduced incidence of peel cracking is associated with increased epidermal cell density, which is maintained until fruit maturation. Presently, the expression of genes that have been previously reported to be associated with epidermal cell patterning and cuticle formation, or cracking resistance, was monitored in the peel during fruit development and following GA4+7 and BA treatment. For most of the genes whose expression is naturally upregulated during fruit development, the early GA4+7 and BA treatment maintained or further increased the high expression level in the mature peel. Where the expression of a gene was downregulated during development, no change was detected in the treated mature peel. Gene-networking analysis supported the interaction between gene clusters of cell-wall synthesis, cuticle formation and GA signaling. Overall, the data suggested that the GA4+7 and BA treatment did not modify developmental cues, but promoted or enhanced the innate developmental program.

Potato root system development and factors that determine its architecture.

The potato root system is often characterized as shallow and inefficient, with poor ability to extract water and minerals from the soil. Potato root system architecture (RSA) refers to its 3-dimensional structure as determined by adventitious root (AR) growth and branching through lateral roots (LR). Understanding how the root system develops holds potential to increase plant yield and optimize agricultural land use. Root system development was monitored in greenhouse-grown potato while a root-on-a-plate assay was developed to explore factors that affect AR and LR development. Expression study of LR-related genes was conducted. Transgenic plants carrying DR5:GFP and CycB1:GUS reporter genes were used to monitor auxin signaling and cell division during root primordia formation, respectively. Maximum root development occurred mainly during the 6-week post seed-tuber planting and slowed during the onset of tuberization. AR and LR development was coordinated - a positive correlation was found between the length of AR and LR and between LR length and number. The expression of LR-related genes was higher in LR than in AR. High nitrate levels reduced LR number and length, however ablation of root-cap by high temperature (33°C) or cutting resulted with enhanced formation of LR. Growth conditions affect AR and LR development in potato, determining the final architecture of its root system. The overall results indicate that LR formation in potato follows similar pattern as in model plants, facilitating study and manipulation of its RSA to improve soil exploitation and yield.

Oligosaccharide mass profiling of nutritionally important Salicornia brachiata, an extreme halophyte.

Salicornia brachiata is an extreme halophyte, growing opulently in salt marshes and considered as potential alternative crop for seawater agriculture. Salicornia seeds are rich in protein and tender-shoots are eaten as salad greens. Total cell wall carbohydrate was isolated from seedlings, digested with ß-glucanase enzyme cocktail and oligosaccharide mass profiling (OLIMP) was performed by using MALDI TOF-TOF mass spectroscopy. Salicornia OLIMP is represented by characteristic mass peaks m/z 477.3297-2094.4363. MS spectra exhibit xyloglucan oligosaccharide building blocks, dominated by XXXG (1084.9927 m/z). Characteristic mass peaks (m/z) of xyloglucan oligomers (XXG, XXFG, XLXG and XLFG) were also detected, which showed resemblance to the mass profile of highly nutritious plant soybean. The present study is the first report on OLIMP for any edible halophyte, so far. OLIMP supports use of Salicornia as a potential source of dietary supplementation. Further linkage-analysis is required to get the structural information of oligosaccharides.

Developing transgenic Jatropha using the SbNHX1 gene from an extreme halophyte for cultivation in saline wasteland.

Jatropha is an important second-generation biofuel plant. Salinity is a major factor adversely impacting the growth and yield of several plants including Jatropha. SbNHX1 is a vacuolar Na⁺/H⁺ antiporter gene that compartmentalises excess Na⁺ ions into the vacuole and maintains ion homeostasis. We have previously cloned and characterised the SbNHX1 gene from an extreme halophyte, Salicornia brachiata. Transgenic plants of Jatropha curcas with the SbNHX1 gene were developed using microprojectile bombardment mediated transformation. Integration of the transgene was confirmed by PCR and Rt-PCR and the copy number was determined by real time qPCR. The present study of engineering salt tolerance in Jatropha is the first report to date. Salt tolerance of the transgenic lines JL2, JL8 and JL19 was confirmed by leaf senescence assay, chlorophyll estimation, plant growth, ion content, electrolyte leakage and malondialdehyde (MDA) content analysis. Transgenic lines showed better salt tolerance than WT up to 200 mM NaCl. Imparting salt tolerance to Jatropha using the SbNHX1 gene may open up the possibility of cultivating it in marginal salty land, releasing arable land presently under Jatropha cultivation for agriculture purposes. Apart from this, transgenic Jatropha can be cultivated with brackish water, opening up the possibility of sustainable cultivation of this biofuel plant in salty coastal areas.