Differential absorption of cadmium and zinc by tobacco plants: Role of apoplastic pathway.

Cadmium (Cd) contamination presents a significant challenge in global agriculture. This study explores the efficacy of chemical induction, specifically using sodium chloride (NaCl), to limit Cd uptake in tobacco (Nicotiana tabacum) and assesses its impact on essential divalent metal ions (DMIs). We conducted a comprehensive analysis encompassing ion absorption, root histology, and biochemistry to understand the influence of this method. Our results revealed that NaCl induction led to a notable 30 % decrease in Cd absorption, while maintaining minimal impact on zinc (Zn) uptake. Intriguingly, the absence of essential DMIs, such as calcium (Ca), magnesium (Mg), and Zn, was found to diminish the plant's capacity to absorb Cd. Furthermore, moderate NaCl induction resulted in an increased diameter of the root stele and enhanced lignin content, indicating a restriction of Cd absorption through the apoplastic pathway. Conversely, a compensatory absorption mechanism via the symplastic pathway appeared to be activated in the absence of essential elements. These findings highlight the potential of chemical induction as a strategy to mitigate agricultural Cd risks, offering insights into the complex interplay between plant ion transport pathways and metal uptake regulation.

Characterization of salt tolerant wheat genotypes by using morpho-physiological, biochemical, and molecular analysis.

Food security is facing a major threat from salinity and there is a need to develop salt tolerant crop varieties to ensure that the demand for food from the world's increasing population is met. Salinity mostly occurs in arid and semi-arid regions. It may cause many adverse physiological effects on plants, i.e., toxic ion accumulation, disturbed osmotic potential, and decreased crop yield. The present study aimed to investigate the morphological, physiological, biochemical, and genetic parameters of wheat genotypes under salt stress. Six wheat genotypes were screened for salt tolerance at the seedling and maturity stage. Seeds were sown at 0 and 150 mM of salinity level. Biochemical traits, i.e., shoot/root fresh and dry weight, chlorophyll a/b and total chlorophyll contents, shoot nitrogen, shoot phosphorus, proline, and carbohydrates were measured. Wheat genotypes showed a significant increase in free amino acids, shoot nitrogen, and total soluble proteins under saline conditions. Higher Na+/K+ ratio and free amino acids were estimated under 150 mM NaCl treatment in Pasban-90 and found to be the most salt-tolerant genotype. By contrast, reduced proline, total chlorophyll, and Na+/K+ ratio were found in Kohistan-97 marking it to be sensitive to stress. Expression analysis of HKTs genes was performed to validate the results of two contrasting genotypes. The differential expression of HKT2; 1 and HKT2; 3 explained the tissue and genotype specific epigenetic variations. Our findings indicated that these selected genotypes can be further used for molecular studies to find out QTLs/genes related to salinity. This suggests that, in contrasting wheat genotypes, there is a differentially induced defense response to salt stress, indicating a functional correlation between salt stress tolerance and differential expression pattern in wheat.

Using CRISPR-Cas9 Technology to Eliminate Xyloglucan in Tobacco Cell Walls and Change the Uptake and Translocation of Inorganic Arsenic.

Xyloglucan is a quantitatively major polysaccharide in the primary cell walls of flowering plants and has been reported to affect plants' ability to tolerate toxic elements. However, it is not known if altering the amounts of xyloglucan in the wall influences the uptake and translocation of inorganic arsenic (As). Here, we identified two Nicotiana tabacum genes that encode xyloglucan-specific xylosyltransferases (XXT), which we named NtXXT1 and NtXXT2. We used CRISPR-Cas9 technology to generate ntxxt1, ntxxt2, and ntxxt1/2 mutant tobacco plants to determine if preventing xyloglucan synthesis affects plant growth and their ability to accumulate As. We show that NtXXT1 and NtXXT2 are required for xyloglucan biosynthesis because no discernible amounts of xyloglucan were present in the cell walls of the ntxxt1/2 double mutant. The tobacco double mutant (ntxxt1/2) and the corresponding Arabidopsis mutant (atxxt1/2) do not have severe growth defects but do have a short root hair phenotype and a slow growth rate. This phenotype is rescued by overexpressing NtXXT1 or NtXXT2 in atxxt1/2. Growing ntxxt mutants in the presence of AsIII or AsV showed that the absence of cell wall xyloglucan affects the accumulation and translocation of As. Most notably, root retention of As increased substantially and the amounts of As translocated to the shoots decreased in ntxxt1/2. Our results suggest that xyloglucan-deficient plants provide a strategy for the phytoremediation of As contaminated soils.

Induction of Semi-Dwarf Trait to a Three Pistil Tall Mutant Through Breeding With Increased Grain Numbers in Wheat.

Multi-ovary wheat (three pistil) is a unique germplasm for the seed production of hybrid wheat. The purpose of the present study was to transfer the multi-ovary trait to semi-dwarf plants to increase the production of grains in wheat crops. Therefore, tall, semi-dwarf, and dwarf plants were crossed with plants with the three-pistil trait. A three-pistil tall plant was used as the female parent, while tall (Synthetic hexaploid), semi-dwarf, and dwarf plants were used as male parents. F1 and F2 progenies with parents were planted in 2015-16 using RCBD. The outcome of the crosses showed that multi-ovary tall plants gave significant difference for all five traits (days to maturity, plant height, number of seeds per spike, grain weight per spike, and grain yield per unit area) in both generations. The greatest number of grains per spike and grain yield per unit area were obtained from the cross of three-pistil tall and dwarf parent (P1/P6) in the F1 and F2 generations. The cross also resulted in a significant reduction in height (96 cm). Further heterosis studies conducted with crosses between three-pistil tall and dwarf parent (P1/P6) showed the greatest heterosis and heterobeltiosis for the number of grains per spike (60.0 and 26.19%, respectively) and grain yield per m2 (27.68 and 2.85%, respectively). In the case of grain weight per spike, the heterosis value was also positive and significant (17.7). Meanwhile, for other traits, their values were negative for heterosis and heterobeltiosis. High numbers of grains and grain weight were found to be associated with positive heterobeltiosis and in turn the grain yield per m2, but plant height and maturity had negative affirmation with heterobeltiosis. Heterosis studies also indicated the dominant gene action for the three-pistil trait. Thus, the study clearly signified that grain yield can be increased by using the multi-ovary genotype with the semi-dwarf height. This new germplasm will be helpful for breeders to increase the production of wheat crops in the southern climate of Pakistan.

Identification and characterization of pectin related gene NbGAE6 through virus-induced gene silencing in Nicotiana benthamiana.

Virus-induced gene silencing (VIGS) is a transient based reverse genetic tool used to elucidate the function of novel gene in N. benthamiana. In current study, 14 UDP-D-glucuronate 4-epimerase (GAE) family members were identified and their gene structure, phylogeny and expression pattern were analyzed. VIGS system was optimized for the functional characterization of NbGAE6 homologous genes in N. benthamiana. Whilst the GAE family is well-known for the interconversion of UDP-D-GlcA and UDP-D-GalA during pectin synthesis. Our results revealed that the downregulation of these genes significantly reduced the amount of GalA in the homogalacturunan which is the major component of pectin found in primary cell wall. Biphenyl assay and high performance liquid chromatography analysis (HPLC) depicted that the level of 'GalA' monosaccharide reduced to 40-51% in VIGS plants as compared to the wild type plants. Moreover, qRT-PCR also confirmed the downregulation of the NbGAE6 mRNA in VIGS plants. In all, this is the first comprehensive study of the optimization of VIGS system for the provision of rapid silencing of GAE family members in N. benthamiana, eliminating the need of stable transformants.

Research Progress on Plant RING-Finger Proteins.

E3 ubiquitin ligases are the most expanded components of the ubiquitin proteasome system (UPS). They mediate the recognition of substrates and later transfer the ubiquitin (Ub) of the system. Really Interesting New Gene (RING) finger proteins characterized by the RING domain, which contains 40-60 residues, are thought to be E3 ubiquitin ligase. RING-finger proteins play significant roles in plant growth, stress resistance, and signal transduction. In this study, we mainly describe the structural characteristics, classifications, and subcellular localizations of RING-finger proteins, as well the physiological processes of RING-finger proteins in plant growth and development. We also summarize the functions of plant RING-finger proteins in plant stress resistance. Finally, further research on plant RING-finger proteins is suggested, thereby establishing a strong foundation for the future study of plant RING-finger proteins.

Genome-Wide Identification, Characterization and Expression Patterns of the Pectin Methylesterase Inhibitor Genes in Sorghum bicolor.

Cell walls are basically complex with dynamic structures that are being involved in several growth and developmental processes, as well as responses to environmental stresses and the defense mechanism. Pectin is secreted into the cell wall in a highly methylesterified form. It is able to perform function after the de-methylesterification by pectin methylesterase (PME). Whereas, the pectin methylesterase inhibitor (PMEI) plays a key role in plant cell wall modification through inhibiting the PME activity. It provides pectin with different levels of degree of methylesterification to affect the cell wall structures and properties. The PME activity was analyzed in six tissues of Sorghum bicolor, and found a high level in the leaf and leaf sheath. PMEI families have been identified in many plant species. Here, a total of 55 pectin methylesterase inhibitor genes (PMEIs) were identified from S. bicolor whole genome, a more detailed annotation of this crop plant as compared to the previous study. Chromosomal localization, gene structures and sequence characterization of the PMEI family were analyzed. Moreover, cis-acting elements analysis revealed that each PMEI gene was regulated by both internal and environmental factors. The expression patterns of each PMEI gene were also clustered according to expression pattern analyzed in 47 tissues under different developmental stages. Furthermore, some SbPMEIs were induced when treated with hormonal and abiotic stress. Taken together, these results laid a strong foundation for further study of the functions of SbPMEIs and pectin modification during plant growth and stress responses of cereal.

Tubby-like Protein 2 regulates homogalacturonan biosynthesis in Arabidopsis seed coat mucilage.

KEY MESSAGE: A possible transcription factor TLP2 was identified to be involved in the regulation of HG biosynthesis in Arabidopsis seed mucilage. TLP2 can translocate into nucleus from plasma membrane by interacting with NF-YC3. The discovery of TLP2 gene function can further fulfill the regulatory network of pectin biosynthesis in Arabidopsis thaliana. Arabidopsis seed coat mucilage is an excellent model system to study the biosynthesis, function and regulation of pectin. Rhamnogalacturonan I (RG-I) and homogalacturonan (HG) are the major polysaccharides constituent of the Arabidopsis seed coat mucilage. Here, we identified a Tubby-like gene, Tubby-like protein 2 (TLP2), which was up-regulated in developing siliques when mucilage began to be produced. Ruthenium red (RR) staining of the seeds showed defective mucilage of tlp2-1 mutant after vigorous shaking compared to wild type (WT). Monosaccharide composition analysis revealed that the amount of total sugars and galacturonic acid (GalA) decreased significantly in the adherent mucilage (AM) of tlp2-1 mutant. Immunolabelling and dot immunoblotting analysis showed that unesterified HG decreased in the tlp2-1 mutant. Furthermore, TLP2 can translocate into nucleus by interacting with Nuclear Factor Y subunit C3 (NF-YC3) to function as a transcription factor. RNA-sequence and transactivation assays revealed that TLP2 could activate UDP-glucose 4-epimerase 1 (UGE1). In all, it is concluded that TLP2 could regulate the biosynthesis of HG possibly through the positive activation of UGE1.

Progress in Optimization of Agrobacterium-Mediated Transformation in Sorghum (Sorghum bicolor).

This review archives the achievements made in the last two decades and presents a brief outline of some significant factors influencing the Agrobacterium-mediated transformation of Sorghum bicolor. Recently, progress in successful transformation has been made for this particular monocot crop through direct DNA delivery method and indirect method via Agrobacterium. However, lower transformation rate still proved to be a bottleneck in genetic modification of sorghum. An efficient Agrobacterium transformation system could be attained by optimizing the preliminary assays, comprising of explant source, growth media, antibiotics, Agrobacterium strains and agro-infection response of callus. The selection of competent strains for genetic transformation is also one of the key factors of consideration. Successful transformation is highly dependent on genome configuration of selected cultivar, where non-tannin genotype proved the best suited. Immature embryos from the field source have higher inherent adaptation chances than that of the greenhouse source. A higher concentration of Agrobacterium may damage the explant source. Utilization of anti-necrotic treatments and optimized tissue culture timeframe are the adequate strategies to lower down the effect of phenolic compounds. Appropriate selection of culture media vessels at different stages of tissue culture may also assist in a constructive manner. In conclusion, some aspects such as culture environment with medium composition, explant sources, and genotypes play an indispensable role in successful Agrobacterium-mediated sorghum transformation system.