Cloning of acyl-acyl carrier protein (ACP) thioesterase gene from Brassica juncea.

Acyl-acyl carrier protein (ACP) thioesterase is the chain-length-determining enzyme in de novo biosynthesis of plant fatty acids. For cloning the gene encoding acyl-ACP thioesterase from Brassica juncea, genomic DNA was used as a template to amplify a 0.7 kb thioesterase fragment in a PCR with the primers designed from the known sequences available in the GenBank. This 0.7 kb fragment was used as a probe to study the expression of the gene in developing seeds and also to screen a genomic library of B. juncea constructed in lambdaEMBL-3 to get the full length of the gene. A 4.0 kb BamHI fragment containing the full gene was finally cloned in a plasmid vector from a recombinant phage clone lambda5.12 after a series of screening, sub-cloning and Southern hybridization.

Effect of water stress on proline content and transcript levels in Lathyrus sativus.

Response of Lathyrus sativus plants to water stress showed that ABA responsive genes such as PLE 25, TAS 14 and RAB 17 are synthesized constitutively, the levels of which decline gradually with increase in water stress or ABA levels. Proline accumulation was highest in leaves (65-fold) followed by stem (56-fold), root (38-fold) and marginal increase in etiolated seedlings. Proline increase was also observed in plant parts not exposed to light.

Differential carbon monoxide sensitivity of cytochrome C oxidase in the leaves of c(3) and c(4) plants.

CO sensitivity of cytochrome a(3) in the leaves of a number of C(3) and C(4) plants was monitored by the nitrate reductase assay under differing CO to O(2) ratios. All the C(3) plants were relatively insensitive to CO and required a high CO to O(2) ratio of 40 to promote significant nitrate reductase activity. However, when treated with 2 millimolar 2,4-dinitrophenol, these leaves readily responded to CO even at low CO to O(2) ratios of 10 or less. On the other hand, the leaves of all C(4) plants tested, belonging to the three subgroups, were highly sensitive to CO, even at CO to O(2) ratios of 5 or less. In these leaves, the uncoupler was without any effect, probably because the mitochondria, either from mesophyll or bundle sheath cells or both, lacked tight respiratory control.

Regulation of hordein synthesis in barley high lysine mutant Notch-2.

Genomic DNA isolated from barley cv. NP 113 and its high lysine mutant Notch-2, and restricted with different restriction enzymes was hybridized with B1 and C-hordein DNA probes. Similar Southern hybridization patterns were observed between NP 113 and Notch-2. Dot blot hybridization analysis of RNA isolated at different developmental stages and from different tissues of seed showed temporal as well as tissue specific expression. The results obtained indicate that regulation at the level of transcription/post transcription may be responsible for lower accumulation of hordein in mutant Notch-2.

Construction of chickpea genomic library.

For construction of chickpea genomic library, DNA was isolated, purified on CsCl gradient and size fractionated into 15-20 Kb fragments after restriction with Sau 3A. These fragments were ligated to phage lambda (EMBL-3) vector and the recombinant molecules packaged in vitro into viable phage particles. The recombinant phages were obtained as phages on a P2 lysogen of E. coli (Spi- selection) and amplified to establish a permanent library. This is the first report of the construction of chickpea genomic library.

Amino acid compositions of different protein fractions in developing grains of NP 113 barley and its high lysine Notch-2 mutant.

The percent distributions of protein fractions namely albumin + globulin, prolamine and glutelin were studied in developing grains of NP 113 barley and its high lysine mutant Notch-2. During development the percentage of albumin + globulin fraction decreased in NP 113, while those of prolamine and glutelin remained unchanged. The increase in prolamine was substantial from 24 to 31DAA. In Notch-2 the trend followed by albumin + globulin and prolamine was like that in NP 113, while the glutelin fraction showed an increase as compared to 10 DAA. The percent of albumin + globulin was slightly higher in Notch-2 as compared to NP 113. The absolute amount (mg/grain) of all the protein fractions increased during development in both NP 113 and its mutant Notch-2. During the grain development the prolamine content was substantially lower in the mutant than in the parent NP 113. The albumin + globulin content per endosperm was in general also higher in NP 113 than Notch-2. Amino acid analysis of the protein fractions did not reveal significant changes in lysine between NP 113 and Notch-2. Thus, the improvement in lysine in the mutant is primarily due to reduced synthesis of the prolamine fraction and not due to an increase in lysine in the mutant hordein fraction. Part of the improvement in lysine may also be due to increase in the percentage of albumin + globulin fractions which is lysine rich.

Soluble proteins and isoenzymes from seeds of diverse male steriles of sorghum, (Sorghum bicolor (L.) Moench).

A comparison of soluble protein, esterase, GDH and ADH isoenzyme patterns in seeds of different steriles, maintainers and restorer lines exhibited similarities as well as differences. Soluble protein patterns from sterile and maintainer lines differed both qualitatively and quantitatively. Based on the esterase patterns, male steriles with different cytoplasms could be separated into three groups (i) Ck 60A and B; Nagpur A and B, (ii) M 35-1A and 1 B, M 31-2A and 2B, (iii) G1A and B, VZM2A and 2B. Each group could further be differentiated on the basis of minor differences in esterase isoenzyme patterns within each group. ADH and GDH patterns in general were similar in both sterile and maintainer lines.

Amino acids in anthers of Milo and in cytoplasmic genetic male sterile sorghums (Sorghum bicolor L. Moench) of Indian origin.

Amino acid composition of proteins from anthers of milo and Indian origin male steriles were determined. Comparison of amino acid between A and B lines showed lower contents of histidine, threonine, glutamic acid, glycine, leucine and phenylalanine and higher contents of alanine, serine, proline and tyrosine in line A compared to line B. Alanine content in anthers of A lines was more than two fold higher than that in the anthers from B lines. Marked differences in amino acid composition of anthers of A and B lines are suggestive of their involvement in male sterility. Cytoplasmic male steriles of Indian origin M35-1A and M31-2A showed greater similarity but differed from milo, VZM2A and B.

Peroxidase activities in relation to plant height and grain weight in bread wheat (Triticum aestivum L.).

The relationship of peroxidase activity with plant height and grain weight has been studied in seven different varieties of bread wheat belonging to diverse genotypes, and their F1 crosses. The association between plant height and peroxidase activity was highly significant and negative. Based on the similarity index values of peroxidase isoenzymes, the seven wheat genotypes could be classified into two groups: the first group consisting of triple and quadruple dwarf varieties and the other of tall, single and double dwarf. A negative correlation between peroxidase activity and grain weight was also observed. However, the results of this study indicate a possibility of developing a dwarf plant type with low peroxidase activity and well filled grains.