Global incidence of wheat rusts and powdery mildew during 1969–2010 and durability of resistance of winter wheat variety Bezostaya 1

Disease incidence and severity was studied for winter wheat variety Bezostaya 1 and susceptible checks based on data from international nurseries from 1969 to 2010 and from 51 countries across major winter wheat production regions totalling 1,047 reports. The frequency of leaf rust and stripe rust occurrence was stable over time with marked increases in severity in 2001–2010 especially in Europe and Central and West Asia. Substantial global reductions in stem rust occurrence were recorded and attributed primarily to use of resistance genes although the recent emergence of race Ug99 makes wheat more vulnerable. The occurrence of powdery mildew remained globally stable over time. It was the most important foliar disease in Western and Southern Europe, where the frequency was very high for all time periods coupled with slight increases in severity during 2001–2010. The durable resistance of variety Bezostaya 1 to all four diseases was demonstrated in the study using comparisons of disease severities of Bezostaya 1 and the most susceptible entries. The Lr34/Yr18/Pm38 pleiotrophic set possessed by Bezostaya 1 is currently an important target for selection because it is now amenable to molecular selection. Increased use of genes like Lr34 combined with strategies to minimize cultivation of extremely susceptible varieties will contribute to long term maintenance of low and non-damaging disease levels. The durable disease resistance of Bezostaya 1, combined with its adaptability and good end-use quality, was a significant reason for its huge impact in agriculture over the last 50?years.     

New sources of resistance to Fusarium wilt and sterility mosaic disease in a mini-core collection of pigeonpea germplasm

Fusarium wilt (FW) and Sterility mosaic disease (SMD) are important biotic constraints to pigeonpea production worldwide. Host plant resistance is the most durable and economical way to manage these diseases. A pigeonpea mini-core collection consisting of 146 germplasm accessions developed from a core collection of 1290 accessions from 53 countries was evaluated to identify sources of resistance to FW and SMD under artificial field epiphytotic conditions during 2007–08 and 2008–09 crop seasons. Resistant sources identified in the field were confirmed in the greenhouse using a root dip screening technique for FW and a leaf stapling technique for SMD. Six accessions (originated from India and Italy were found resistant to FW (<10% mean disease incidence). High level of resistance to SMD was found in 24 accessions (mean incidence <10%). These SMD resistant accessions originated from India, Italy, Kenya, Nepal, Nigeria, Philippines and United Kingdom. Combined resistance to FW and SMD was found in five accessions (ICPs 6739, 8860, 11015, 13304 and 14819). These diverse accessions that are resistant to FW or SMD will be useful to the pigeonpea resistance breeding program.     

Comparative analyses of spot blotch and tan spot epidemics on wheat under optimum and late sowing period in South Asia

In South Asia, foliar blight of wheat (Triticum aestivum L.) is a disease complex caused by Cochliobolus sativus (the spot blotch pathogen) and Pyrenophora tritici-repentis (the tan spot pathogen) which can reduce yields by >30?%. Little is known about the effects of wheat genotypes and their planting time on foliar blight epidemics. Field experiments were conducted to determine the infection potential and epidemic development of C. sativus and P. tritici-repentis on two susceptible (Sonalika and BL1473) and two tolerant (NL750 and Milan/Shanghai-7) wheat genotypes under optimum (November 26) and late (December 11 and December 26) planting conditions. The dynamics of airborne conidia were studied using air samplers. The highest aerial concentrations of conidia and disease incidence of both pathogens on all four wheat genotypes were detected during the first 3?weeks of March under both optimum and late seeding conditions in both years. Compared to optimum sowing time (i.e. November 26), wheat genotypes had higher disease severity when planted late on December 11 and 26. The disease complex reduced grain yield by 20.5, 27.2 and 37.3?% for November 26, December 11 and December 26 plantings, respectively in 2004. The corresponding differences were 17.7, 23.6 and 30.2?% in 2005. The findings of this study provide valuable information on the epidemiology of spot blotch and tan spot, which could help in developing strategies for managing these diseases in South Asian region through the selection of suitable genotypes and planting dates.     

Effects of fertilizers and plant strengtheners on the susceptibility of tomatoes to single and mixed isolates of Phytophthora infestans

Three organic fertilizers, Horn meal, i.e., BioFeed Basis and Bio-ILSA, and three plant strengtheners, i.e., Alfalfa extract, PEN and QUALITY were compared to chemical fertilizer (NPK) application and the synthetic inducer BABA (DL-3-amino-butyric acid) for reactions of leaf disks of six different tomato cultivars to three isolates of Phytophthora infestans. Late blight severity was significantly reduced on plants fertilized with Bio-ILSA and BioFeed Basis compared to plants fertilized with Horn meal and chemical fertilizer. There were no interactions between fertilizers and isolates or between fertilizers and cultivars. All plant strengtheners significantly reduced susceptibility of all tomato cultivars. However, effects were isolate and tomato cultivar specific. The reductions in area under the disease progress curve compared to the water control for the different tomato cultivars and isolates were 23–78%, 21–77%, 17–66%, and 37–100% for Alfalfa extract, PEN, QUALITY and BABA, respectively. Similar, but somewhat smaller reductions were observed for sporulation capacity. The plant strengtheners were more effective in controlling disease on plants challenged with isolate mixtures than on plants challenged with single isolates. BABA performed significantly better than the plant strengtheners in 65% (i.e., 34 out of 54) of the tested cultivar-isolate combinations, when single isolates were used. When two isolates were mixed, the percentage was reduced to 45% (25 out of 54 cases) and to 33%, (6 out of 18 cases) with a mixture of three isolates.     

Characterization of biofumigated Ralstonia solanacearum cells using micro-Raman spectroscopy and electron microscopy

Essential oils of palmarosa, lemongrass, and eucalyptus have shown promise as biofumigants for control of the bacterial wilt disease of edible ginger (Zingiber officinale) caused by Ralstonia solanacearum race 4 in previous potting medium studies. Biochemical changes in R. solanacearum cells were evaluated with micro-Raman spectroscopy following treatment with essential oils at different concentrations (0.04, 0.07, and 0.14% [vol/vol] of culture medium) and changes in cell structure were observed using electron microscopy. All treatments except palmarosa oil at 0.04% caused significant reductions in levels of amino acids, purine and pyrimidine bases of nucleic acids, carbohydrates, and lipids, as indicated by significant reduction in Raman peak heights at 621, 1,003, and 1,031 inverse centimeters (cm(-1)) (phenylalanine); 643, 827, 852, 1,158, and 1,172 cm(-1) (tyrosine); 758 cm(-1) (tryptophan); 725, 782, 1,337, and 1,578 cm(-1) (adenine, cytosine plus uracil, adenine, and adenine plus guanine, respectively); 1,097 cm(-1) (carbohydrates); and 1,127, 1,450, and 2,932 cm(-1) (lipids) compared with untreated controls. Lemongrass oil treatments were the most effective in degrading cellular components. Scanning electron microscopy of palmarosa and lemongrass-oil-treated cells showed rupture of cell walls and cell debris but no degradation was noted for eucalyptus-oil-treated cells. Palmarosa- and lemongrass-oil-treated cells were positively stained with uranyl acetate when viewed by transmission electron microscopy whereas controls and eucalyptus-oil-treated cells were negatively stained, indicating that the cell membranes were intact. The viability of eucalyptus-oil-treated cells was confirmed by cell culture following treatment. Micro-Raman spectroscopy is a powerful tool which can be further employed to better understand effects of fumigants and other bactericides on bacterial cells.     

Expression and Characterization of Constitutive Heat Shock Protein 70.1 (HSPA‐1A) Gene in In Vitro Produced and In Vivo‐Derived Buffalo (Bubalus bubalis) Embryos

Cells are blessed with a group of stress protector molecules known as heat shock proteins (HSPs), amongst them HSP70, encoded by HSPA‐1A gene, is most abundant and highly conserved protein. Variety of stresses hampers the developmental competence of embryos under in vivo and in vitro conditions. Present work was designed to study the quantitative expression of HSPA‐1A mRNA in immature oocytes (IMO), matured oocytes (MO), in vitro produced (IVP) and in vivo‐derived (IVD) buffalo embryos to assess the level of stress to which embryos are exposed under in vivo and in vitro culture conditions. Further, HSPA‐1A gene sequence was analysed to determine its homology with other mammalian sequences. The mRNA expression analysis was carried out on 72 oocytes (40 IMO; 32 MO), 76 IVP and 55 IVD buffalo embryos. Expression of HSPA‐1A was found in oocytes and throughout the developmental stages of embryos examined irrespective of the embryo source; however, higher (p < 0.05) expression was observed in 8–16 cell, morula and blastocyst stages of IVP embryos as compared to IVD embryos. Phylogenetic analysis of bubaline HSPA‐1A revealed that it shares 91–98% identity with other mammalian sequences. It can be concluded that higher level of HSPA‐1A mRNA in IVP embryos in comparison with in vivo‐derived embryos is an indicator of cellular stress in IVP system. This study suggests need for further optimization of in vitro culture system in which HSPA‐1A gene could be used as a stress biomarker during pre‐implantation development.     

Oxidant/antioxidant balance, minerals status and apoptosis in peripheral blood of dogs naturally infected with Dirofilaria immitis

The present study was aimed to examine lipid peroxides, antioxidants, zinc, copper, iron panels and apoptosis in peripheral blood of dogs with clinical dirofilariosis. In comparison to healthy control, erythrocytic lipid peroxides level and superoxide dismutase activity were significantly (P<0.01) higher in diseased dogs. However, level of reduced glutathione and activities of glutathione peroxidase, glutathione-S-transferase and catalase were significantly (P<0.01) lower in diseased dogs. Blood zinc, copper and iron levels in dogs with dirofilariosis were significantly (P<0.01) lower than the healthy control. The percent apoptotic peripheral leukocytes were significantly higher in dogs with dirofilariosis in comparison with healthy control. From the present study, it is concluded that significant alterations in oxidant/antioxidant balance, mineral status and rate of apoptosis in peripheral blood may be implicated in the pathogenesis of clinical canine dirofilariosis.     

AFLP fingerprinting of tartary buckwheat accessions (Fagopyrum tataricum) displaying rutin content variation

In light of the economic importance of buckwheat as well as existence of enormous accessions of Fagopyrum species in the Himalayan regions of India, the characterization of tartary buckwheat for rutin content variation vis-à-vis DNA fingerprinting was undertaken so as to identify fingerprint profiles unique to high rutin content accessions. Rutin content analysis in mature seeds of 195 accessions of Fagopyrum tataricum showed a wide range of variation (6 μg/mg to 30 μg/mg D.W.) with most of the accessions (81%) containing 10-16 μg/mg of rutin followed by 14% accessions with significantly higher rutin content (17 μg/mg to 30 μg/mg) and 5% accessions with low rutin content (≤10 μg/mg). AFLP fingerprinting of 18 accessions having high (≥17 μg/mg) and low rutin content (≤10 μg/mg) with 19 EcoRI/MseI primer combinations yielded 136 polymorphic fragments out of total 907. The hierarchical and model-based cluster analyses of AFLP data strongly suggested that the 18 populations of F. tataricum were clustered into two separate groups. The high and low rutin content accessions were clustered into two separate groups based on AFLP fingerprinting. The AFLP fingerprints associated with high rutin content accessions of F. tataricum are expected to be useful for evaluation, conservation and genetic improvement of buckwheat.     

Purification, characterization and evaluation of insecticidal activity of trypsin inhibitor from Albizia lebbeck seeds

A Bowman-Birk inhibitor with activity against gut proteases of Helicoverpa armigera was extracted in 0.1 M sodium phosphate buffer from defatted seed flour of Albizia lebbeck. It was purified to 29.62 folds with 51.43% recovery using ammonium sulfate precipitation, gel filtration chromatography on Sephadex G-100 column and ion exchange chromatography on DEAE-Sephadex A50. The purified protein had a molecular weight of 12,303 daltons as determined by SDS-PAGE. It was found to be heat stable up to 60°C and had two pH optima of 7.5 and 9.0. The inhibitor exhibited non-competitive pattern of inhibition with a low Ki value of 0.2 μM. The inhibitor was found to be susceptible to varying concentrations of reducing agents like DTT and 2-mercaptoethanol, thereby indicating the role of disulphide bridges in maintaining its three dimensional structure and stability. The purified inhibitor caused mortality and suppressed larval growth of Pieris brassicae larvae. It was also found to be effective against gut trypsin extracted from Spodoptera littoralis. The sequence of the genes encoding for such inhibitors can be determined and the genes expressing protease inhibitors can be used in vegetable crops to confer resistance against insect pests and other plant pathogens.