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Roots from salt-susceptible ICSR-56 (SS) sorghum plants display metaxylem elements with thin cell walls and large diameter. On the other hand, roots with thick, lignified cell walls in the hypodermis and endodermis were noticed in salt-tolerant CSV-15 (ST) sorghum plants. The secondary wall thickness and number of lignified cells in the hypodermis have increased with the treatment of sodium chloride stress to the plants (STN). Lignin distribution in the secondary cell wall of sclerenchymatous cells beneath the lower epidermis was higher in ST leaves compared to the SS genotype. Casparian thickenings with homogenous lignin distribution were observed in STN roots, but inhomogeneous distribution was evident in SS seedlings treated with sodium chloride (SSN). Higher accumulation of K+ and lower Na+ levels were noticed in ST compared to the SS genotype. To identify the differentially expressed genes among SS and ST genotypes, transcriptomic analysis was carried out. Both the genotypes were exposed to 200 mM sodium chloride stress for 24 h and used for analysis. We obtained 70 and 162 differentially expressed genes (DEGs) exclusive to SS and SSN and 112 and 26 DEGs exclusive to ST and STN, respectively. Kyoto Encyclopaedia of Genes and Genomes (KEGG) and Gene Ontology (GO) enrichment analysis unlocked the changes in metabolic pathways in response to salt stress. qRT-PCR was performed to validate 20 DEGs in each SSN and STN sample, which confirms the transcriptomic results. These results surmise that anatomical changes and higher K+/Na+ ratios are essential for mitigating salt stress in sorghum apart from the genes that are differentially up- and downregulated in contrasting genotypes.

The structural changes in the secretory cells are important to understand the ontogeny and nectar secretion process from the nectaries. In this study, we investigated the ultrastructural changes during different developmental/secretion stages of extrafloral nectaries (EFNs) of Pithecellobium dulce. The dense cytoplasm with active biosynthesis mechanisms such as ribosomes, mitochondria, large nucleus, and plastids with accumulated starch grains characterized the pre-secretion stage of young nectariferous cells. During the secretory phase, the cytoplasm showed distinct changes associated with endomembrane transport such as the predominant occurrence of Golgi, secretory vesicles, and ER resulting in the subsequent appearance of secretions in the intercellular and subcuticular spaces. Cell wall loosening following the dissolution of middle lamellae leading to the formation of subcuticular spaces was evident during advanced stages of nectar secretion. The characteristic cytoplasmic and apoplastic changes associated with cell death were noticed during the post-secretory stages. The structural evidence from the present study suggests the occurrence of two modes of secretion (merocrine and holocrine) during the early and late stages of secretion in the EFNs of P. dulce.

Plant growth regulators play a key role in cell wall structure and chemistry of woody plants. Understanding of these regulatory signals is important in advanced research on wood quality improvement in trees. The present study is aimed to investigate the influence of exogenous application of 24-epibrassinolide (EBR) and brassinosteroid inhibitor, brassinazole (BRZ) on wood formation and spatial distribution of cell wall polymers in the xylem tissue of Leucaena leucocephala using light and immuno electron microscopy methods. Brassinazole caused a decrease in cambial activity, xylem differentiation, length and width of fibres, vessel element width and radial extent of xylem suggesting brassinosteroid inhibition has a concomitant impact on cell elongation, expansion and secondary wall deposition. Histochemical studies of 24-epibrassinolide treated plants showed an increase in syringyl lignin content in the xylem cell walls. Fluorescence microscopy and transmission electron microscopy studies revealed the inhomogenous pattern of lignin distribution in the cell corners and middle lamellae region of BRZ treated plants. Immunolocalization studies using LM10 and LM 11 antibodies have shown a drastic change in the micro-distribution pattern of less substituted and highly substituted xylans in the xylem fibres of plants treated with EBR and BRZ. In conclusion, present study demonstrates an important role of brassinosteroid in plant development through regulating xylogenesis and cell wall chemistry in higher plants.

Osmotin like protein ( OLP ) and chitinase ( Chi11 ) belong to pathogenesis-related (PR) class of proteins and are induced during both biotic and abiotic stresses. Transgenic tomato was developed with OLP and Chi11 via in vitro and in planta transformation methods. Transgenes integration and transcript levels were confirmed by multiplex PCR, DNA blot, and multiplex reverse transcriptase PCR. Homozygous T 2 transgenics when evaluated for salt, drought, and fungal stresses showed enhanced tolerance compared to untransformed controls (UC). Transgenics showed enhanced endochitinase activity and root biomass under normal conditions. Transformants also displayed higher proline content, K + , relative water content, chlorophyll fluorescence, total biomass, vascular conductivity, and fruit yield than the UC under stress conditions. Co-immunoprecipitation revealed that Chi11 co-expresses with phosphofructokinase2 (PFK2), which may play a role in enhanced root biomass. qPCR analysis resulted in higher transcript levels of OLP , Chi11, and PFK2 in transgenics as compared to the untransformed controls. Our findings suggest the use of multiples genes to confer multiple stress tolerance for enhanced crop productivity. This work unveils a new molecular player PFK2, which may play a role in enhanced root biomass.

A Na+/H+ antiporter-like protein (NHXLP) was isolated from Sorghum bicolor L. (SbNHXLP) and validated by overexpressing in tomato for salt tolerance. Homozygous T2 transgenic lines when evaluated for salt tolerance, accumulated low Na+ and displayed enhanced salt tolerance compared to wild-type plants (WT). This is consistent with the amiloride binding assay of the protein. Transgenics exhibited higher accumulation of proline, K+, Ca2+, improved cambial conductivity, higher PSII, and antioxidative enzyme activities than WT. Fluorescence imaging results revealed lower Na+ and higher Ca2+ levels in transgenic roots. Co-immunoprecipitation experiments demonstrate that SbNHXLP interacts with a Solanum lycopersicum cation proton antiporter protein2 (SlCHX2). qRT-PCR results showed upregulation of SbNHXLP and SlCHX2 upon treatment with 200 mM NaCl and 100 mM potassium nitrate. SlCHX2 is known to be involved in K+ acquisition, and the interaction between these two proteins might help to accumulate more K+ ions, and thus maintain ion homeostasis. These results strongly suggest that plasma membrane bound SbNHXLP involves in Na+ exclusion, maintains ion homeostasis in transgenics in comparison with WT and alleviates NaCl stress.

A Na+/H+ antiporter-like protein (NHXLP) was isolated from Sorghum bicolor L. (SbNHXLP) and validated by overexpressing in tomato for salt tolerance. Homozygous T2 transgenic lines when evaluated for salt tolerance, accumulated low Na+ and displayed enhanced salt tolerance compared to wild-type plants (WT). This is consistent with the amiloride binding assay of the protein. Transgenics exhibited higher accumulation of proline, K+, Ca2+, improved cambial conductivity, higher PSII, and antioxidative enzyme activities than WT. Fluorescence imaging results revealed lower Na+ and higher Ca2+ levels in transgenic roots. Co-immunoprecipitation experiments demonstrate that SbNHXLP interacts with a Solanum lycopersicum cation proton antiporter protein2 (SlCHX2). qRT-PCR results showed upregulation of SbNHXLP and SlCHX2 upon treatment with 200 mM NaCl and 100 mM potassium nitrate. SlCHX2 is known to be involved in K+ acquisition, and the interaction between these two proteins might help to accumulate more K+ ions, and thus maintain ion homeostasis. These results strongly suggest that plasma membrane bound SbNHXLP involves in Na+ exclusion, maintains ion homeostasis in transgenics in comparison with WT and alleviates NaCl stress.

Sequential changes occurring in cell walls during expansion, secondary wall (SW) deposition and lignification have been studied in the differentiating xylem elements of Holoptelea integrifolia using transmission electron microscopy. The PATAg staining revealed that loosening of the cell wall starts at the cell corner middle lamella (CCML) and spreads to radial and tangential walls in the zone of cell expansion (EZ). Lignification started at the CCML region between vessels and associated parenchyma during the final stages of S2 layer formation. The S2 layer in the vessel appeared as two sublayers,an inner one and outer one.The contact ray cells showed SW deposition soon after axial paratracheal parenchyma had completed it, whereas noncontact ray cells underwent SW deposition and lignification following apotracheal parenchyma cells. The paratracheal and apotracheal parenchyma cells differed noticeably in terms of proportion of SW layers and lignin distribution pattern. Fibres were found to be the last xylem elements to complete SW deposition and lignification with differential polymerization of cell wall polysaccharides. It appears that the SW deposition started much earlier in the middle region of the fibres while their tips were still undergoing elongation. In homogeneous lignin distribution was noticed in the CCML region of fibres.

The anatomy of the secondary xylem and distribution pattern of gelatinous fibres (G-fibres) have been studied in the developing and heavy fruit bearing mature peduncles of Kigelia pinnata and Couroupita guianensis. The peduncle in both the plants developed reaction xylem as a result of growth stresses caused by development of large fruits. In Couroupita peduncles which are originally horizontal, G-fibre distribution was unilateral and similar to that of typical tension wood whereas the hanging Kigelia peduncles have uniformly distributed gelatinous fibres throughout the xylem. The tension xylem severity was higher in the basal region and decreased towards the terminal region of the current year’s peduncle but after fruit development a drastic increase in tension wood severity was observed in the terminal region. In the Kigelia peduncles, tension wood severity in terms of G-layer proportion to lignified wall was found to be less than in Couroupita. The abundance of vessels decreased with high frequency of gelatinous fibres in Couroupita. The peduncle of Kigelia is characterized by high vessel frequency, thin normal fibre walls, and thick outer walls with thin gelatinous layer in tension wood fibres. Dimensional variations were also noticed in the mechanical and conducting elements varying with tension wood severity.