Regeneration and micrografting of lentil shoots

Summary A protocol for regeneration and micrografting of shoots of lentil (Lens culinaris Medik) was developed. Multiple shoots (4–5) were regenerated from cotyledonary node explants on Murashige and Skoog (MS) medium containing 8.8 μM 6-benzylaminopurine. In vitro regenerated shoots were micrografted on rootstocks with 96% efficiency. The successful grafts were transplanted to pots in Redi-earth^TM, hardened off and were grown to maturity with 100% success. The success of the micrografting was independent of the nature and concentration of growth regulator used in shoot initiation medium and the time period for induction of shoots. The protocol was successful with several cultivars of lentil. The advantages of micrografting over in vitro rooting are discussed.     

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Effect of ischemia-reperfusion injury on the morphology of peroxisomes

We have previously demonstrated that ischemic injury changed the density of peroxisomes into two distinct peaks, one with a normal density (1.21 g/cm³; Peak I) and a second peak with a lighter density (1. 14 g/cm³; Peak II).We studied the peroxisomes from both peaks under the Electron microscope. Examination of peak I following ischemia showed loss of matrix proteins and damaged limiting membranes with leakage of DAB positive material in direct proportion to the duration of ischemia. Upon reperfusion of the ischemic liver Peak I showed more severe damage to the organelle. These observations clearly demonstrated that ischemia reperfusion injury causes structural damage to peroxisomes. Interestingly ultrastructural examination of Peak II following ischemia showed evidence of perisomal proliferation with budding of existing peroxisomes and the presence of micro peroxisomes (changes similar to those noted under conditions leading to perisomal proliferation). However, peak II following reperfusion showed only damaged organelle. These observations underline the importance of peroxisomes in the response of the cell to ischemia-reperfusion injury.     

Lactose repressor hinge domain independently binds DNA

The short 8–10 amino acid “hinge” sequence in lactose repressor (LacI), present in other LacI/GalR family members, links DNA and inducer‐binding domains. Structural studies of full‐length or truncated LacI‐operator DNA complexes demonstrate insertion of the dimeric helical “hinge” structure at the center of the operator sequence. This association bends the DNA ～40° and aligns flanking semi‐symmetric DNA sites for optimal contact by the N‐terminal helix‐turn‐helix (HtH) sequences within each dimer. In contrast, the hinge region remains unfolded when bound to nonspecific DNA sequences. To determine ability of the hinge helix alone to mediate DNA binding, we examined (i) binding of LacI variants with deletion of residues 1–50 to remove the HtH DNA binding domain or residues 1–58 to remove both HtH and hinge domains and (ii) binding of a synthetic peptide corresponding to the hinge sequence with a Val52Cys substitution that allows reversible dimer formation via a disulfide linkage. Binding affinity for DNA is orders of magnitude lower in the absence of the helix‐turn‐helix domain with its highly positive charge. LacI missing residues 1–50 binds to DNA with ～4‐fold greater affinity for operator than for nonspecific sequences with minimal impact of inducer presence; in contrast, LacI missing residues 1–58 exhibits no detectable affinity for DNA. In oxidized form, the dimeric hinge peptide alone binds to O1 and nonspecific DNA with similarly small difference in affinity; reduction to monomer diminished binding to both O1 and nonspecific targets. These results comport with recent reports regarding LacI hinge interaction with DNA sequences.     

Indigenous salt-tolerant rhizobacterium <Emphasis Type="Italic">Pantoea dispersa</Emphasis> (PSB3) reduces sodium uptake and mitigates the effects of salt stress on growth and yield of chickpea

Salt stress has multiple damaging effects on plants including physiological damage, reduced growth, and productivity. Plant growth-promoting rhizobacteria (PGPR) are one of the valuable options to mitigate the negative effects of this stress. In the present study, native bacteria from chickpea’s rhizosphere were isolated, and checked for their salt tolerance and plant growth-promoting attributes (phosphate (P) solubilization, siderophores, indole-3-acetic acid (IAA) production, and 1-aminocyclopropane-1-carboxylate (ACC) deaminase production). One isolate, subsequently identified as Pantoea dispersa, showed appreciable production of IAA (218.3 µg/ml) and siderophores (60.33% SU), P-solubilization (3.64 µg/ml) and ACC deaminase activity (207.45 nmol/mg/h) in the presence of 150 mM NaCl, under laboratory conditions. Salt stress in uninoculated chickpea (GPF2 cultivar) plants induced high accumulation of Na⁺ ions (3.86 mg g^?1 dw) in the leaves, along with significant reduction in K⁺ uptake, membrane integrity, chlorophyll concentration, and leaf water content, thus resulting in impaired growth of the plant and yield (pods and seeds) in a salt concentration-dependent manner. The damage due to salt stress was restored significantly in plants inoculated with P. dispersa. A significant improvement in biomass (32–34%), pods number (31–34.5%), seeds number (32–35.7%), pods weight (30–32.6%), and seeds weight (27–35%) per plant occurred in salt stress-affected plants, which was associated with significant reduction in Na⁺ uptake, reduced membrane damage, significantly improved leaf water content, chlorophyll content, and K⁺ uptake. This study suggests for the first time that native P. dispersa strain PSB3 can be used to alleviate the negative effects of salt stress on chickpea plants and holds the potential to be used as a biofertilizer.