小麦条锈菌PsNCS1基因的克隆及转录表达特征

摘要：【目的】克隆小麦条锈菌神经钙离子感应蛋白基因PsNCS1,分析其在病菌不同发育时期的表达水平。【方法】利用文库筛选和RT-PCR技术克隆PsNCS1的cDNA序列,采用生物信息学技术预测分析该基因编码蛋白的保守结构域及基本特性,构建系统发育树;运用实时荧光定量RT-PCR技术分析PsNCS1在病菌不同生长发育时期的表达水平。【结果】PsNCS1全长cDNA为1007 bp（GenBank登录号GU134621）,开放阅读框为573 bp,编码190个氨基酸,分子量为22.17 kDa, 等电点为4.     

Association of some plant defense enzyme activities with systemic resistance to early leaf blight and leaf spot induced in tomato plants by azoxystrobin and Pseudomonas fluorescens

Abstract

Azoxystrobin at three different concentrations, namely, 31.25, 62.50 and 125 g a.i. ha^?1 mancozeb (1 kg ha^?1) and Pseudomonas fluorescens (10 kg ha^?1) were evaluated for their efficacy in inducing defense enzymes in tomato against Alternaria solani and Septoria lycopersici. The activity of defense enzymes peroxidase (PO), polyphenol oxidase (PPO), phenylalanine ammonia lyase (PAL), β-1, 3 glucanase, chitinase, catalase and defense-inducing chemicals (total phenols) was found to be increased in azoxystrobin and P. fluorescens-treated tomato plants. The activity of these defense enzymes and chemicals was higher in azoxystrobin (125 g a.i. ha^?1) and P. fluorescens-treated tomato plants challenge inoculated with the pathogens compared to other treatments. Increased expression of specific isoforms of PO and PPO was also observed due to ISR induction.     

Virulence variation of Puccinia striiformis Westend. f. sp. tritici in Pakistan

Yellow rust populations of Pakistan were characterised for their virulence pathotypes/races and pathogenetic variation using seedling evaluation of differential genotypes under glasshouse conditions in Murree (6000 feet above sea level). Differential genotypes comprised a world set, an European set, near isogenic lines and the universally susceptible bread wheat cultivar “Morocco”. Over the two-year study a total of 18 race groups were identified. Out of these 18 race groups, several (68E0, 64E0, 66E0, 70E0, 6E0, 71E0, 6E0, 2E0, 67E0, and 68E16) were found previously. The new race group 70E32 found probably evolved because of mutation from the previously existing 70E16. Virulence frequencies of yellow rust (Yr) resistance genes were also determined on near isogenic lines. The highest virulence frequencies (%) were found for Yr7 (88%), Yr9 (57%), Yr18 (70%), and Yr24 (69%). Virulence frequencies were low for Yr 1 (4%), Yr5 (7%), Yr10 (10%) and Yr15 (4%). Our studies indicated that virulence existed for almost all yr genes, necessitating regular monitoring of the yellow rust populations and intensifying efforts to identify new sources of resistance to this pathogen.     

Methyl jasmonate stimulates polyamines biosynthesisand resistance against leaf rust in wheat plants

Abstract

Leaf rust of wheat (Triticum aestivum L.), incited by Puccinia recondita ex Desm. f. sp. tritici Eriks, is one of the most important wheat diseases in Egypt. Methyl jasmonate (MJ) is a potential plant elicitor which induces a wide range of chemical and anatomical defense reactions in conifers and might be used to increase systemic resistance against biotic damage. In the greenhouse, different concentrations of MJ (10, 20 and 30 mM) were applied as seed soaking plus foliar spray or only as foliar spray to control leaf rust and induction of secondary compound production in leaves of wheat plants. Foliar spray was applied after 30 and 50 days of sowing. Results indicated that all concentrations and treatments reduced the severity of rust disease caused by P. recondita f. sp. tritici in wheat leaves during 45 days of inoculations. Disease incidence was decreased significantly in MJ-treated plants as seed soaking plus foliar spray with 20 and 30 mM when compared to 10 mM MJ or control plants. The study revealed that, with increasing concentrations of MJ, the secondary metabolites were greatly increased. Endogenous levels of both free and conjugated putrescine, spermidine and spermine increased in response to the elicitor. Activities of polyamine biosynthetic enzymes of ornithine decarboxylase (ODC) and polyamine oxidase (PAO) displayed up to threefold increases relative to untreated control. Moreover, significant increases in activities of plant defense-related protein, enzymes as peroxidase and chitinase as well as free and conjugated phenols contents were recorded in treated plants compared with untreated and infected plants. Furthermore, MJ treatment increased the chlorophyll-a, chlorophyll-b and carotenoids pigments contents, the higher increase was obtained with combined treatment between seeds soaking plus foliar spray at 20 and 30 mM of MJ. Under field conditions, three concentrations of MJ, i.e. 10, 20 and 30 mM as combined treatment between seeds soaking plus foliar spray or only as foliar spray were applied to study their effect against rust disease. Foliar spray was applied after 30 and 80 days of sowing. Results showed that the high reduction in disease severity was obtained with combined treatments between seeds soaking plus foliar spray with MJ at 20 and 30 mM compared with other treatments and control. At the same time, all treatments increased the growth and grain yield of wheat plants. It could be suggested that combination treatment between seeds soaking plus foliar spray with methyl jasmonate might be used commercially for controlling rust disease of wheat plants under field conditions.     

Function and evolution of allelic variations of Sr13 conferring resistance to stem rust in tetraploid wheat (Triticum turgidum L.)

The resistance gene Sr13 is one of the most important genes in durum wheat for controlling stem rust caused by Puccinia graminis f. sp. tritici (Pgt). The Sr13 functional gene CNL13 has haplotypes R1, R2 and R3. The R1/R3 and R2 haplotypes were originally designated as alleles Sr13a and Sr13b, respectively. To detect additional Sr13 alleles, we developed Kompetitive allele specific PCR (KASP™) marker KASPSr13 and four semi-thermal asymmetric reverse PCR markers, rwgsnp37–rwgsnp40, based on the CNL13 sequence. These markers were shown to detect R1, R2 and R3 haplotypes in a panel of diverse tetraploid wheat accessions. We also observed the presence of Sr13 in durum line CAT-A1, although it lacked any of the known haplotypes. Sequence analysis revealed that CNL13 of CAT-A1 differed from the susceptible haplotype S1 by a single nucleotide (C2200T) in the leucine-rich repeat region and differed from the other three R haplotypes by one or two additional nucleotides, confirming that CAT-A1 carries a new (R4) haplotype. Stem rust tests on the monogenic, transgenic and mutant lines showed that R1 differed from R3 in its susceptibility to races TCMJC and THTSC, whereas R4 differed from all other haplotypes for susceptibility to TTKSK, TPPKC and TCCJC. Based on these differences, we designate the R1, R3 and R4 haplotypes as alleles Sr13a, Sr13c and Sr13d, respectively. This study indicates that Sr13d may be the primitive functional allele originating from the S1 haplotype via a point mutation, with the other three R alleles probably being derived from Sr13d through one or two additional point mutations.     

Field tests of bacteria and soil-applied fungicides as control agents for take-all in winter wheat

Putative biological and chemical treatments for controlling take-all were used in each of three consecutive years at two locations where winter wheat crops were grown in naturally-infested fields. The chemical treatments more often decreased take-all than the biological treatments, but no treatment consistently and significantly decreased take-all, nor did any cause a significant increase in yield. An isolate of Bacillus cereus var. mycoides and one of B. pumilis, applied as soil drenches in autumn or spring, or in the seed furrows, were usually ineffective. Of the few significant effects on disease, half were associated with increases and half with decreases, and most occurred in April and did not persist to late June. Two strains of Pseudomonas pluorescens applied to the seed were ineffective. The fungicide benomyl, applied as a drench in autumn and spring at 20 kg/ha was ineffective, while nuarimol, applied as a drench in autumn at 2 kg/ha was sometimes effective. Nuarimol incorporated into the seed bed at 2 kg/ha was the most effective treatment. In analyses using a functional relationship model for data from treated and untreated plots 12% of 176 data sets for biological treatments, 38% of 96 data sets for chemical treatments and 81% of 16 data sets for combined treatments showed increasing efficiency of the treatment with increasing disease intensity. These findings also demonstrate an additional advantage of the experimental design, namely that treatments are tested at different disease intensity levels within fields.     

The use of the area under the disease-progress curve (AUDPC) to assess quantitative disease resistance in crop cultivars

Calculation of the area under the disease-progress curve (AUDPC) as a measure of quantitative disease resistance entails repeated disease assessments. For typical sigmoid disease-progress curves, repeated assessments may be unnecessary. A mathematical procedure is derived for estimating the AUDPC from two data points. A field trial with ten cultivars with and without the gene Yr18 for resistance to stripe rust were inoculated with two pathotypes of Puccinia striiformis (from the cultivars Karamu and Oroua) and assessed for the percentage leaf area infected seven or eight times during the growing season. The AUDPCs were calculated directly from data and estimated from the described equation. Calculated values were plotted and ranked against estimated values, and excellent correspondence was obtained (Spearman’s Rank Correlation in the Karamu trial= 0.9879 and the Oroua trial=0.9515). Therefore, an estimation of the AUDPC from two data points provides an equivalent amount of information as from repeated assessments. Received: 23 September 1999 / Accepted: 14 April 2000