Artificial neural network modeling for deciphering the in vitro induced salt stress tolerance in chickpea (Cicer arietinum L).

Salt stress is one of the most critical abiotic stresses having significant contribution in global agriculture production. Chickpea is sensitive to salt stress at various growth stages and a better knowledge of salt tolerance in chickpea would enable breeding of salt tolerant varieties. During present investigation, in vitro screening of desi chickpea by continuous exposure of seeds to NaCl-containing medium was performed. NaCl was applied in the MS medium at the rate of 6.25, 12.50, 25, 50, 75, 100, and 125 mM. Different germination indices and growth indices of roots and shoots were recorded. Mean germination (%) of roots and shoots ranged from 52.08 to 100%, and 41.67-100%, respectively. The mean germination time (MGT) of roots and shoots ranged from 2.40 to 4.78 d and 3.23-7.05 d. The coefficient of variation of the germination time (CVt) was recorded as 20.91-53.43% for roots, and 14.53-44.17% for shoots. The mean germination rate (MR) of roots was better than shoots. The uncertainty (U) values were tabulated as 0.43-1.59 (roots) and 0.92-2.33 (shoots). The synchronization index (Z) reflected the negative impact of elevated salinity levels on both root and shoot emergence. Application of NaCl exerted a negative impact on all growth indices compared to control and decreased gradually with elevated NaCl concentration. Results on salt tolerance index (STI) also revealed the reduced STI with elevated NaCl concentration and STI of roots was less than shoot. Elemental analysis revealed more Na and Cl accumulation with respective elevated NaCl concentrations. The In vitro growth parameters and STI values validated and predicted by multilayer perceptron (MLP) model revealed the relatively high R 2 values of all growth indices and STI. Findings of this study will be helpful to broaden the understanding about the salinity tolerance level of desi chickpea seeds under in vitro conditions using various germination indices and seedling growth indices. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-023-01282-z.

Genetic transformation of common beans (Phaseolus vulgaris L.) through Agrobacterium tumefaciens carrying Cry1Ab gene.

BACKGROUND: Seed beetles are one of the most important causes of yield loss in bean production. It is essential to develop resistant varieties in the fight against these pests. Agrobacterium-based gene transformation is the most widely used breeding method worldwide to develop insect-resistant varieties. METHODS AND RESULTS: Embryonic axes and plumule explants were obtained from Agrobacterium tumefciens treated mature zygotic embryos of low and high raw protein-based common bean cultivars Akman 98 and Karacasehir 90. Agrobacterium tumefaciens contained a synthetic Bacillus thuringiensis insecticidal crystal protein gene (Bt Cry1Ab) controlled by the 35S promoter and NOS terminator sequences. The transformation event was genotype and explant dependent. The plumule explants could not withstand kanamycin-based selection pressure and died. It was possible to get two transgenic plants using embryonic axis explants of low protein cultivar Akman 98. These results were validated using GUS analysis, PCR, RT-PCR, bioassay analysis, and ELISA test from the samples taken from T0 and T1 generations. Bioassay tests showed that these plants were protected from the damage of legume seed insects (Bruchus spp.). CONCLUSIONS: The results are very encouraging and may help in producing better transgenic common bean germplasm leading to safe agriculture and reducing environmental pollutions.

Artificial intelligence-based approaches to evaluate and optimize phytoremediation potential of in vitro regenerated aquatic macrophyte Ceratophyllum demersum L.

Water bodies or aquatic ecosystem are susceptible to heavy metal accumulation and can adversely affect the environment and human health especially in underdeveloped nations. Phytoremediation techniques of water bodies using aquatic plants or macrophytes are well established and are recognized as eco-friendly world over. Phytoremediation of heavy metals and other pollutants in aquatic environments can be achieved by using Ceratophyllum demersum L. - a well-known floating macrophyte. In vitro regenerated plants of C. demersum (7.5 g/L) were exposed to 24, 72, and 120 h to 0, 0.5, 1.0, 2.0, and 4.0 mg/L of cadmium (CdSO4·8H2O) in water. Results revealed significantly different relationship in terms of Cd in water, Cd uptake by plants, bioconcentration factor (BCF), and Cd removal (%) from water. The study showed that Cd uptake by plants and BCF values increased significantly with exposure time. The highest BCF value (3776.50) was recorded for plant samples exposed to 2 mg/L Cd for 72 h. Application of all Cd concentrations and various exposure duration yielded Cd removal (%) between the ranges of 93.8 and 98.7%. These results were predicted through artificial intelligence-based models, namely, random forest (RF), extreme gradient boosting (XGBoost), and multilayer perceptron (MLP). The tested models predicted the results accurately, and the attained results were further validated via three different performance metrics. The optimal regression coefficient (R2) for the models was recorded as 0.7970 (Cd water, mg/L), 0.9661 (Cd plants, mg/kg), 0.9797 bioconcentration factor (BCF), and 0.9996 (Cd removal, %), respectively. These achieved results suggest that in vitro regenerated C. demersum can be efficaciously used for phytoremediation of Cd-contaminated aquatic environments. Likewise, the proposed modeling of phytoremediation studies can further be employed more comprehensively in future studies aimed at data prediction and optimization.

Current trends and challenges in the synthesis and applications of chitosan-based nanocomposites for plants: A review.

Chitosan, a low-cost and multipurpose polymer with numerous desired physicochemical and biological properties has been tested for various applications in agriculture, pharmacy, and biomedicine industries. The availability of functional groups along the backbone makes chitosan readily available for other polymers and metal ions to form bio-nanocomposites. Different types of chitosan-based nanocomposites have been designed and tested for the enhancement of chitosan efficiency and ultimately widening the application areas of chitosan in plants. These nanocomposites serve different purposes such as eliciting plant's defence systems against different threats (pathogen attack), antimicrobial agent against bacteria, fungi and viruses, enhancement of nutrient uptake by plants, control release of micro/macronutrients, fungicides and herbicides. In this review, an extensive outlook has been provided (mainly in the last five years) to recent trends and advances in the fabrication and application of chitosan-based composites. Finally, current challenges and future development opportunities of chitosan-based nanocomposites for plants are discussed.

Rediscovering the Potential of Multifaceted Orphan Legume Grasspea- a Sustainable Resource With High Nutritional Values.

The genus Lathyrus consists of more than 184 herbaceous annual and perennial species suitable for multifaceted sustainable food and feed production system in the arid and semi-arid regions of the world. The grasspea is a promising source of protein nutrition. However, its potential is not being utilized fully due to the presence of neurotoxin content (ß-N-oxalyl-l-α, ß diaminopropionic acid, ß-ODAP), a causal agent of non-reversible lower limbs paralysis. The high protein contents in seeds and leaves with ~90% digestibility make it sustainable super food to beat protein malnutrition in future. Therefore, it is desired to breed new grasspea cultivars with low ß-ODAP contents. Limited research has been carried out to date about this feature. A draft genome sequence of grasspea has been recently published that is expected to play a vital role in breeding and identifying the genes responsible for biosynthesis pathway of ß-ODAP contents in grasspea. Efforts to increase awareness about the importance of genus Lathyrus and detoxify ß-ODAP in grasspea are desired and are in progress. Presently, in South Asia, systematic and dedicated efforts to support the farmers in the grasspea growing regions by disseminating low ß-ODAP varieties has resulted in a considerable improvement in reducing the incidence of neurolathyrism. It is expected that the situation will improve further by mainstreaming grasspea cultivation by implementing different approaches such as the development and use of low ß-ODAP varieties, strengthening government policies and improved detox methods. The present review provides insight into the multifaceted characteristics of sustainable nutritious grasspea in the global and Indian perspective.

Understanding the role of phytohormones in cotton fiber development through omic approaches; recent advances and future directions.

Cotton is among the most important fiber crops for the textile-based industry, thanks to its cellulose-rich mature fibers. The fiber initiation and elongation are one of the best models for deciphering mechanisms of single-cell differentiation and growth, that also target of fiber development programs. During the last couple of decades, high yielding omics approaches (genomics, transcriptomics, and proteomics), have helped in the identification of several genes and gene products involved in fiber development along with functional relationship to phytohormones. For example, MYB transcription factor family and Sus gene family have been evidenced by controlling cotton fiber initiation. Most importantly, the biosynthesis, responses, and transporting of phytohormones is documented to participate in the initiation of cotton fibers. Herein, in this review, the reliable genetic evidence by manipulating the above genes in cotton have been summarized to describe the relationships among key phytohormones, transcription factors, proteins, and downstream fiber growth-related genes such as Sus. The effect of other important factors such as ROS, fatty acid metabolism, and actin (globular multi-functional proteins) over fiber development has also been discussed. The challenges and deficiencies in the research of cotton fiber development have been mentioned along with a future perspective to discover new crucial genes using multiple omics analysis.

Chitosan-based delivery systems for plants: A brief overview of recent advances and future directions.

Chitosan has been termed as the most well-known among biopolymers, receiving widespread attention from researchers in various fields mainly, agriculture, food, and health. Chitosan is a deacetylated derivative of chitin, mainly isolated from waste shells of the phylum Arthropoda after their consumption as food. Chitosan molecules can be easily modified for adsorption and slow release of plant growth regulators, herbicides, pesticides, and fertilizers, etc. Chitosan as a carrier and control release matrix that offers many benefits including; protection of biomolecules from harsh environmental conditions such as pH, light, temperatures and prolonged release of active ingredients from its matrix consequently protecting the plant's cells from the hazardous effects of burst release. In the current review, tends to discuss the recent advances in the area of chitosan application as a control release system. Also, future recommendations will be made in light of current advancements and major gaps.

Current advancements in chitosan-based film production for food technology; A review.

Chitosan is obtained from chitin, which could be considered to be the most abundant polymer after cellulose. Owing to these properties, chitosan alone or chitosan-based composite film production is attaining huge attention in terms of applications from researchers and industrialists coming from divergent fields. To enhance the biological (mainly antimicrobial and antioxidant) and physiological (mainly mechanical, thermal and barrier) attributes of the chitosan-based films, a vast medley of plant extracts and supporting polymers has been blended into chitosan films. Considering the up to date literature reports based on chitosan film production and applications, it can be stated that still, the research ratio is low in this field. Chitosan blend/composite films with specific properties (superhydrophobicity, excellent mechanical strength, acceptable barrier properties) can be produced only for specific applications in food technology. In the current review, we tried to summarize the advancements made in the last 5-7 years in the field of chitosan film technology for its application in the food industry.

Production of novel chia-mucilage nanocomposite films with starch nanocrystals; An inclusive biological and physicochemical perspective.

In the current study, chia mucilage composite films with starch nanocrystals (3% and 6%) were produced. The films were analyzed physicochemically (FT-IR, AFM, TGA, DSC), mechanically (Tensile strength and contact angle) and biologically (antimicrobial, antioxidant and cytotoxicity) properties. The incorporation of starch nanocrystals was confirmed through FT-IR spectra showing broad OH peak and CO stretching and shift in NH bending vibrations to the lower wave number. Starch nanocrystals enhanced (control 287.23 °C, film with 3% SNC 286.91 °C and film with 6% mucilage 289.41 °C) the thermal properties of the composite films. The Young Modulus of the film showed an increase after the incorporation of starch nanocrystals due to the strong interaction between mucilage and nanocrystals. On the other hand, the overall hydrophobicity of mucilage composite film decreased due to the hydrophilic nature of cornstarch nanocrystals. MTT assay for cell proliferation revealed significant inhibition of cancer cell (HepG2) lines and exhibits a very low inhibition of epithelial cell line (Vero). Starch nanocrystals enhanced the antibacterial and antioxidant (threefold increase compare to control) properties of mucilage composite films. Mucilage-SNC composite films could be a good therapeutic gain for control and directed drug delivery, food packaging, food coating.

Interaction between seed size and NaCl on germination and early seedling growth of some Turkish cultivars of chickpea (Cicer arietinum L.).

Chickpea is an important food legume crop of Turkey and is largely grown for human consumption on low moisture or salt-affected soils. The objective of the study was to find the effects of NaCl stress at electrical conductivities of 4.5, 8.6, 12.7 and 16.3 dS/m and seed sizes (7, 8 and 9 mm) on germination and early seedling growth of three popular chickpea cultivars (AKN-97, Gokce and Uzunlu-99). Mean frequency of germination, germination time, germination index, root length, shoot length and seedling fresh weight showed seed size-dependent responses of cultivars to salt stress. In general, small seeds germinated and grew more rapidly compared to medium and large seeds of the same cultivars against all levels of salt stress, with the best results in cultivar Uzunlu-99. No effect of NaCl treatments was observed on frequency of germination; however, a drastic decrease in early seedling growth was recorded at increased NaCl concentrations. Regression analysis results showed a significantly positive relationship (P<0.01) between seed size and mean germination time, whereas a significantly negative relationship was recorded between seed size and germination index, root length, shoot length. Moreover, linear regression values apparently confirmed that increased seed size in each cultivar affected decreased germination index, root and shoot lengths with enhanced mean germination time. Thus, it was concluded that the use of small seeds could considerably reduce the production costs of chickpea in salt-affected soils.

Factors affecting efficient in vitro micropropagation of Muscari muscarimi Medikus using twin bulb scale.

Endemic Muscari muscarimi Medikus is the most fragrant plant among Muscari species and has a high ornamental potential. The natural populations of M. muscarimi, are severely affected by increased environmental pollution and urbanization. There is a need to develop a micropropagation method that should serve effectively for commercial propagation and conservation. Therefore, the study targeted to set up a strategy for efficient in vitro bulblet regeneration system of M. muscarimi using twin scale bulb explants on 1.0 × MS medium containing 4.44, 8.88, 17.76 µM BAP (6-Benzylaminopurine) plus 2.685, 5.37, 10.74 µM NAA (α-Naphthalene acetic acid). Maximum number of 19 daughter axillary bulblets and 16 daughter adventitious bulblets per twin bulb scale explant was regenerated on 1.0 × MS medium containing 17.76 µM BAP plus 10.74 µM NAA and 17.76 µM BAP plus 2.685 µM NAA respectively. The daughter bulblets regenerated on twin bulb scales on 8 out of 9 regeneration treatment could be easily rooted on 1.0 × MS medium containing 4.9 µM IBA (Indole-3-butyric acid). The daughter bulblets regenerated on 9th treatment (1.0 × MS medium containing 17.76 µM BAP plus 10.74 µM NAA) were transferred to 1.0 × MS medium containing 30 g/l sucrose to break negative carry over effect of this dose of BAP-NAA, where they grew 2-3 roots of variable length. Daughter bulblet diameter was increased by culturing them on 1.0 × MS medium containing 4.44 µM BAP plus 5.37 µM NAA. The results verified that both age and the source of explants had significant effect on regeneration. In another set of experiments, twin scales were obtained from in vitro regenerated daughter bulblets, although they induced bulblets, yet their bulblet regeneration percentage, mean number of bulblets per explant and their diameter were significantly reduced. In vitro regenerated bulblets were acclimatized in growth chamber under ambient conditions of temperature and humidity on peat moss, where they flowered. The study provides important information about selection of suitable micropropagation medium, strategies to improve bulblet diameter and rooting of M. muscarimi which offers a scope for commercial propagation.

miR408 overexpression causes increased drought tolerance in chickpea.

Drought stress limits yield severely in most of the crops. Plants utilize complex gene regulation mechanisms to tolerate water deficiency as well as other abiotic stresses. MicroRNAs (miRNAs) are a class of small non-coding RNAs that are progressively recognized as important regulators of gene expression acting at post-transcriptional level. miR408, conserved in terrestrial plants, targets copper related genes. Although, expression level of miR408 is influenced by various environmental factors including drought stress, the biological action of miR408 is still unclear. To examine the miR408 function upon drought stress in chickpea, transgenic lines overexpressing the miR408 were generated. Induced tolerance was observed in the plants with enhanced miR408 expression upon 17-day water deficiency. Expression levels of miR408 target gene together with seven drought responsive genes were measured using qRT-PCR. Here, the involvement of miR408 in drought stress response has been reported. The overexpression leading plantacyanin transcript repression caused regulation of DREB and other drought responsive genes.

Factors affecting in vitro plant regeneration of the critically endangered Mediterranean knapweed (Centaurea tchihatcheffii Fisch et. Mey).

Habitat destruction has resulted in the extinction of many plant species from the earth, and many more face extinction. Likely, the annual endemic Mediterranean knapweed (Centaurea tchihatcheffii) growing in the Golbasi district of Ankara, Turkey is facing extinction and needs urgent conservation. Plant tissue culture, a potentially useful technique for ex situ multiplication, was used for the restoration of this ill-fated plant through seed germination, micropropagation from stem nodes, and adventitious shoot regeneration from immature zygotic embryos. The seeds were highly dormant and very difficult to germinate. No results were obtained from the micropropagation of stem nodes. However, immature zygotic embryos showed the highest adventitious shoot regeneration on Murashige and Skoog (MS) medium, containing 1 mg l(-1) kinetin and 0.25 mg l(-1) NAA. Regenerated shoots were best rooted on MS medium containing 1 mg l(-1) IBA and transferred to the greenhouse for flowering and seed set. As such, the present work is the first record of in vitro propagation of critically endangered C. tchihatcheffii, using immature zygotic embryos, and is a step forward towards conservation of this indigenous species.