Evaluation of Wheat Polyphenol Oxidase Genes

Polyphenol oxidases (PPOs) present in mature wheat (Triticum aestivum L.) kernels have been implicated in the undesirable darkening of cereal products such as Asian noodles. To accelerate the functional characterization of wheat PPOs and allow the identification of those PPO genes that are primarily involved in food biochemistry, several basic local alignment search tool (BLAST) searches of expressed sequence tag (EST) databases were performed using a known wheat PPO sequence as a search argument; identified ESTs were resequenced and aligned. Results from this study suggest the presence of at least six PPO genes in hexaploid wheat, falling into two clusters with three similar sequences each. Based on the tissues used for cDNA library preparation, three genes (all members of one cluster) are expressed during kernel development and may therefore influence cereal product quality; the remaining three genes (belonging to the second cluster) were isolated from nonkernel cDNA libraries and may not be expressed at high levels during grain development. Discovery of these genes represents an essential first step in the functional characterization of wheat PPOs.     

Hypodermosis in cattle translocated to Tamil Nadu from Punjab

Tropical Animal Health and Production - Fifteen apparently healthy Kandari cross-bred cattle aged about 4 years were purchased from Rurki, Patiala district of Punjab by a private dairy...     

Novel role of photoinsensitive alleles in adaptation of soybean [Glycine max (L.) Merr.] to rainfed short growing seasons of lower latitudes

Soybean is a temperate photosensitive crop but has adapted to sub-tropical and tropical countries of lower latitudes also. Photoperiodic and maturity genes confer latitudinal adaptation in this crop. Genotyping of accessions of higher latitudes have shown the role of photoinsensitivity, conferred by recessive photoperiodic alleles (e1/e2/e3/e4), in adaptation of the crop to high latitudes but information is not available for lower latitudinal countries like India. We genotyped and calculated the photosensitivity of 101 cultivated Indian soybean varieties and found that majority of the varieties (86) were photosensitive and had the dominant alleles at these loci. Four genotypic classes (e1-as/E2/E3/E4, E1/e2/e3/E4, E1/e2/E3/E4 and E1/E2/e3/E4) were observed for varieties with recessive alleles. Photoinsensitive alleles at E1 and E2 loci significantly reduced the days to flower, maturity and photosensitivity percentage. Adaptive role of photoperiodic alleles was inferred from breeder seed requirement of these varieties for 35 years. Although the photosensitive class contributed 81% to the total seed requirement the weighted mean contribution of this class (380 Q/year) was far less than that of photoinsensitive class (648 Q/year). Photoinsensitivity is essential for perpetuation of crop in higher latitudes. Present report highlights the novel role of photoinsensitive alleles in adaptation of soybean to rainfed, short growing and sub-tropical conditions of lower latitudes by conferring earliness.

     

Genetic analysis of important antioxidant compounds in cabbage (Brassiaca oleracea var. capitata L.)

The paucity of research on development of antioxidant-rich hybrids in cabbage by utilizing combining ability estimates, gene action, and heterosis motivated us to undertake this study. We made 60 crosses between 5 cytoplasmic male-sterile lines and 12 male-fertile testers during the summer of 2015, as per the line × tester design. The seedlings of all the parents and 60 F₁ crosses, along with three checks, were transplanted during the Rabi (winter) season of 2015–2016 and evaluated using a randomized complete-block design. Combining ability, gene action, and heterosis were determined for different antioxidant compounds. Experimental results revealed that the range of cupric ion reducing antioxidant capacity (CUPRAC) [parents = 1.26–7.33 and hybrids = 0.04–6.54 μmol trolox/g], ferric reducing ability of plasma (FRAP) [parents = 1.65–4.76 and hybrids = 0.16–4.67 μmol trolox/g], β-carotene (parents = 0.44–2.29 and hybrids = 0.04–1.89 μg/100 g), and chlorophyll-a (parents = 0.71–4.08 and hybrids = 0.19–3.08 mg/g f.w.) for hybrids was lower than that of the parents because of outbreeding depression. The parental lines 6A, 208A, 83-5-8, and Sel-5-83-6 were found to be good general combiners for most of the antioxidant compounds studied. Based on the mean performance, specific combining ability effects and heterosis, five hybrid combinations viz., 9A × KIRC-8 for CUPRAC and FRAP; 208A × C-122 for ascorbic acid; 6A × Chhaki-2 for total carotenoids and β-carotene; 831A × Chhaki-2 for chlorophyll-a; and 6A × 83-5-8 for chlorophyll-b and total chlorophyll content, were most promising. The ratio of general combining ability (GCA) and specific combining ability (SCA) variances, i.e., [2 σ²g/(2σ²g + σ²s)], which reflects the relative importance of GCA versus SCA, was less than unity for different antioxidant compounds, which implied that for these traits, nonadditive gene effects were more important than additive effects. The numerical values of range for contribution of lines × testers interaction for different traits (41.47–70.18%) were found to be higher than the individual contribution of lines (11.24–47.22%) and testers (8.31–21.76%). Hence, heterosis could be exploited for developing antioxidant-rich hybrid cabbage.     

Development of a loop-mediated isothermal amplification procedure as a sensitive and rapid method for detection of 'candidatus Liberibacter solanacearum' in potatoes and Psyllids

This study reports the development of a loop-mediated isothermal amplification procedure (LAMP) for polymerase chain reaction (PCR)-based detection of 'Candidatus Liberibacter solanacearum', the bacterial causal agent of potato zebra chip (ZC) disease. The 16S rDNA gene of 'Ca. Liberibacter solanacearum' was used to design a set of six primers for LAMP PCR detection of the bacterial pathogen in potato plants and the psyllid vector. The advantage of the LAMP method is that it does not require a thermocycler for amplification or agarose gel electrophoresis for resolution. Positive LAMP results can be visualized directly as a precipitate. The LAMP strategy reported here reliably detected 'Ca. Liberibacter solanacearum' and the closely related species 'Ca. Liberibacter asiaticus', the causative agent of huanglongbing disease of citrus, in plant DNA extracts. Although not as sensitive as quantitative real-time PCR, LAMP detection was equivalent to conventional PCR in tests of ZC-infected potato plants from the field. Thus, the LAMP method shows strong promise as a reliable, rapid, and cost-effective method of detecting 'Ca. Liberibacter' pathogens in psyllids and field-grown potato plants and tubers.     

Studies on a mixed bacterial culture from soil which degrades the herbicide linuron

A stable mixed bacterial culture which degrades the herbicide linuron was isolated from soil by enrichment with linuron in a liquid mineral medium. Radio-respirometry studies showed that the culture mineralised linuron completely. No intermediate degradation products were detected in the medium. The culture was able to utilise linuron as a source of both nitrogen and carbon and was also able to degrade the related herbicides monolinuron and chlorbromuron and the possible intermediate degradation products of linuron: 3,4-dichlorophenyl-l-methylurea, 3,4-dichlorophenylurea and 3,4-dichloroaniline. The culture was unable to degrade the 1,1-dimethyl substituted ureas monuron, diuron or metoxuron. The culture contained Gram-negative aerobic rods, and Gram-positive aerobic non-spore-forming rods and cocco-bacilli. Of 124 isolates from the mixed culture, none degraded linuron in pure culture, indicating that a consortium of organisms is involved. Further investigation suggested that Pseudomonas spp. were important components of the population responsible for degradation.     

Development of a CAPS marker for the badh2.7 allele in Sri Lankan fragrant rice (Oryza sativa)

Fragrance in rice is caused by mutations in the badh2 (betaine aldehyde dehydrogenase) gene. It was previously reported that exons 1, 2, 7, 10, 13 and 14 of badh2 are hot spots for various mutations leading to fragrance in most aromatic rice. This study was carried out to sequence the 14th exon of badh2 gene of Sri Lankan aromatic rice varieties that lack the badh2.1 allele. The aims of the study were to predict the aberrant protein structure and to develop a functional DNA marker. In view of this, we sequenced the 14th exon of four traditional aromatic accessions and compared with a published sequence. Four accessions contained a nucleotide ‘G’ insertion in the 14th exon. This novel mutation can be classified as the badh2.7 allele. The predicted three‐dimensional protein structure of the mutant shows loss of part of the oligomerization and coenzyme binding domains, a change that is predicted to result in fragrance. A CAPS‐based novel marker, Bad2.7CAPS, was developed to identify varieties possessing this badh2.7 allele, and it can be utilized in rice breeding programmes.