Carbohydrate, Amino acid, Phenolic and Mineral Nutrient Contents of Pepper Plants in Relation to Age-Related Resistance to Phytophthora capsici

The relationship between age-related resistance of peper plants to Phytophthora capsici and contents of carbohydrates, amino acids, phenolics and mineral nutrients in pepper stems was studied using two pepper cultivars, Hanbyul (susceptible) and Kingkun (resistant). With increasing age of pepper plants, the two cultivars, which differ in their susceptibility to Phytophthora blight, became gradually resistant to the disease. The cultivar Kingkun distinctly showed the age-related resistance to Phytophthora blight at the second branch stage. The weight of dry matter in healthy stems of pepper plants at the second branch stage was twice that at the six leaf stage. The resistant cultivar Kingkun contained lower levels of fructose, glucose and sucrose in stems than the susceptible cultivar Hanbyul at the different developmental stages. No consistent differences between the developmental stages of the plants were recognized with regard to their glucose content. However, the contents of fructose and sucrose in the cultivar Hanbyul greatly increased at the second branch stage. The levels of inositol reduced in both pepper cultivars during plant development. In view of the fact that there were only slight changes in the amount of total amino acids, it seems unlikely that there is a relationship between the amino acid metabolism and the retardation of Phytophthora infection during plant development. The amounts of total phenolic compounds in pepper stems were relatively low at the later growth stages of the plants and also in the resistant cultivar Kingkun. The contents of macroelemental nutrients such as nitrogen, phosphorus, potassium, calcium and magnesium were drastically reduced in pepper stems at the later plant growth stage. No significant differences between the cultivars or the plant growth stages were found in the silicon and microelemental nutrients such as sodium, iron, zinc and manganese. These results suggest that the expression of age-related resistance of pepper plants may be due to the morphological and nutritional changes in tissues of pepper stems during ageing, i.e. the pronounced increase in weight of dry matter, the significant decrease in amounts of mineral nutrients such as nitrogen, phosphorus, potassium, calcium and magnesium, and the tow contents of fructose, glucose and sucrose in the stem tissues.     

Induction of defence response against blister blight by calcium chloride in tea

Tea, the second most consumed beverage after water in the world, is produced from the processed tender leaves of tea plants (Camellia sinensis). Production of tea is hindered by various biotic and abiotic stresses. Among the biotic factors, blister blight disease caused by an obligate fungal pathogen, Exobasidium vexans Massee, is a serious problem to the tea industry. The present study was to evaluate the efficacy of elicitor calcium chloride (CaCl₂) in inducing resistance in tea plants against blister blight disease. During the pick time of blister incidence at Darjeeling tea garden, the application of CaCl₂ at a concentration of 1% resulted in disease inhibition around 71% over the control set. Treatment also resulted in the induction of defence enzymes like peroxidase, polyphenol oxidase, phenylalanine ammonia lyase, β-1,3-glucanase and higher phenol accumulation. Furthermore, the increase in defence molecules also correlated with increase in nitric oxide (NO) generation, a potent defence molecule in plant defence. The result suggests that CaCl₂ can used as a potential elicitor in the integrated disease management in organic tea cultivation.     

Biochemical and molecular studies on the resistance mechanisms in tea [<Emphasis Type="Italic">Camellia sinensis</Emphasis> (L.) O. Kuntze] against blister blight disease

Tea (Camellia sinensis) plantations are exposed to biotic and abiotic stresses. Among the biotic factors, blister blight (BB), caused by Exobasidium vexans, affects the quality and quantity of the product and demands high fungicide application. A long term solution for disease resistance would require the knowledge of the basic molecular and biochemical changes occurring in plant as an attempt to resist the pathogen and limit the spread of the disease which can further help in developing resistant cultivars using biotechnological tools. Thus, gene expression studies using the cDNA based suppressive subtractive hybridization library, characterization of genes for pathogenesis related (PR) proteins [chitinase (CsCHIT), glucanase (CsGLUC), phenylalanine ammonia lyase (CsPAL)] and genes in flavonoid pathway were accessed in the BB resistant and susceptible cultivars, SA6 and TES34, respectively. Further, biochemical analysis of PR and antioxidant enzymes (POX, APX, SOD) involved in BB resistance have been carried out to investigate the potential molecular and biochemical changes. Various stages of pathogen development had varied impact on PR protein, flavonoid pathway and anti-oxidative enzymes and indicates the possible role of reactive oxygen species, lignins, flavonoids, anthocyanins and other synthesized compounds in acting as antimicrobial/antifungal agents in tea cultivars.     

Study of Agrobacterium-mediated co-transformation of tea for blister blight disease resistance

Journal of Plant Biochemistry and Biotechnology - Blister blight is the most prevalent leaf disease of tea, an economical perennial crop. The disease is caused by a biotrophic fungus, Exobasidium...     

Effects of Fusarium graminearum Metabolites on Wheat Tissue in Relation to Fusarium Head Blight Resistance

Fusarium head blight (FHB) of wheat is caused by Fusarium graminearum which produces many secondary metabolites including the trichothecene mycotoxins deoxynivalenol (vomitoxin) and 3-acetyldeoxynivalenol. Coleoptile tissue segements from 14 spring wheat cultivars were exposed to the F. graminearum metabolites deoxynivalenol, 3-acetyldeoxynivalenol, butenolide (all known mycotoxins), sambucinol, culmorin and dihydroxycalonectrin in a bioassay. The tissue of most cultivars was inhibited, at a concentration of 10^?6M by the trichothecenes tested and up to 10^?3M for the other compounds. Deoxynivalenol and 3-acetyldeoxynivalenol, which affect protein synthesis at the ribosome, are therefore potent phytotoxins in addition to being mycotoxins. The resistance or susceptibility of each cultivar to FHB was established in a field experiment. A comparison of the two sets of data indicated that resistant cultivars could tolerate much higher concentrations of the metabolites tested than susceptible cultivars. Some resistant material can tolerate 10 to 1000 times the concentration of the trichothecenes, compared with susceptible cultivars, with no effect on growth. The data suggest that it may be possible to screen germplasm rapidly for FHB resistance in vitro and a new type of resistance in wheat to this disease is proposed based on the apparent insensitivity to trichothecenes by resistant cultivars, additional to the three types of resistance described in the literature.     

Stable resistance to bacterial blight disease in rice

Ten rice cultivars were evaluated for stable resistance to 18 isolates of Xanthomonas campestris pv. oryzae. Stable performance of the host cultivar was evaluated by regressing the cultivar means on the mean level of pathogenicity of the isolates averaged over all cultivars. A cultivar with low disease incidence, a regression coefficient near or equal to zero (b= 0) and the deviation from regression as small as possible (S²_d= 0) was considered highly stable while that with low disease incidence, b= 1 -0 and S²_d= 0 was considered to possess average stability. Accordingly, cvs TKM-6, CB II, DV 85 and Ramakrishna were considered to possess average stability while IR 20 and Indira were the most stable. Cvs Cemposelak, IR 1545, BJ 1 and IR 8 showed high response to disease incidence with relatively small changes in level of pathogenicity of the isolates and hence were considered most unstable. The regression technique was considered as a valuable tool for identification of stable resistance to bacterial blight disease of rice.     

Comparative proteomic analysis of methyl jasmonate‐induced defense responses in different rice cultivars

Jasmonate is an important endogenous chemical signal that plays a role in modulation of plant defense responses. To understand its mechanisms in regulation of rice resistance against the fungal pathogen Magnaporthe oryzae, comparative phenotype and proteomic analyses were undertaken using two near‐isogenic cultivars with different levels of disease resistance. Methyl‐jasmonate (MeJA) treatment significantly enhanced the resistance against M. oryzae in both cultivars but the treated resistant cultivar maintained a higher level of resistance than the same treated susceptible cultivars. Proteomic analysis revealed 26 and 16 MeJA‐modulated proteins in resistant and susceptible cultivars, respectively, and both cultivars shared a common set of 13 proteins. Cumulatively, a total of 29 unique MeJA‐influenced proteins were identified with many of them known to be associated with plant defense response and ROS accumulation. Consistent with the findings of proteomic analysis, MeJA treatment increased ROS accumulation in both cultivars with the resistant cultivar showing higher levels of ROS production and cell membrane damage than the susceptible cultivar. Taken together, our data add a new insight into the mechanisms of overall MeJA‐induced rice defense response and provide a molecular basis of using MeJA to enhance fungal disease resistance in resistant and susceptible rice cultivars.     

Area under disease progress curve and apparent infection rate of Alternaria blight disease of Indian mustard (Brassica juncea) at different plant age

Plant age has a major influence on the incidence of Alternaria blight disease in Indian mustard crops. Disease progression was monitored twice a week on the two chosen Indian mustard cultivars viz., Varuna and Rohini throughout the season. Severity of blight caused by Alternaria brassicae and Alternaria brassicicola decreased with delay in sowing. Calculation for A-value (Area under disease progress curve – AUDPC) and r-value (apparent infection rate) in crops sown on different dates could identify the speed of progress in the disease on leaves and pods, as the crop does not posses resistance to the pathogen till date. Thus, the probable dates of sowing enabling slow disease progress or the weather conditions coinciding with the different crop phenological stages demarcated the advantageous dates of sowing from the disadvantageous ones. However, cultivar Varuna is more susceptible as compared to the other cultivar Rohini, as apparent infection rate both on leaves and pods was higher in former. Highest per cent disease severity (PDS) for season highly correlated with date of sowing, i.e. delayed date of sowing increased PDS.     

Development of pea bacterial blight caused by Pseudomonas syringae pv. pisi in winter and spring cultivars of combining peas (Pisum sativum) with different sowing dates

Pea bacterial blight occurred by natural infection in a field trial on peas in 1995. Disease development in the winter cultivars Rafale, Frilene and Froidure was compared with that in the spring cultivars Baccara, Conquest and Bohatyr, each sown on six dates in October, November, December, mid-March, late March and April. Disease incidence had reached 100% plants affected in all treatments by mid-July. Disease severity was greater in winter-sown (October, November or December) than in spring-sown peas of each cultivar at each assessment. Significant (P < 0.05) differences in disease severity occurred between cultivars in the winter-sown plots in May and June and the spring cultivars were affected more severely than the winter cultivars. Comparison of areas under the disease progress curves for both disease incidence and severity also showed that the winter-sown peas were more affected by disease than spring-sown peas and that spring cultivars were more severely affected than winter cultivars. Yield was strongly correlated with disease severity. A linear regression model suggested that, for peas sown in October, November or December, a yield loss of 0.5 tha^-1 occurred for each 10% increase in canopy area affected by pea bacterial blight.     

Differences in the Induction of Defence Responses in Resistant and Susceptible Muskmelon Plants Infected with Colletotrichum lagenarium

Defence reactions occurring in resistant (cv. Gankezaomi) and susceptible (cv. Ganmibao) muskmelon leaves were investigated after inoculating with Colletotrichum lagenarium. Lesion restriction in resistant cultivars was associated with the accumulation of hydrogen peroxide (H₂O₂). The activity of antioxidants catalase (CAT) and peroxidase (POD) significantly increased in both cultivars after inoculation, while levels of both CAT and POD activity were significantly higher in the resistant cultivar. Ascorbate peroxidase (APX) increased in both cultivars after inoculation, and level of APX activity was significantly higher in the resistant cultivar. Glutathione reductase (GR) activity significantly increased in both cultivars following inoculation, but was higher in the resistant cultivar, resulting in higher levels of ascorbic acid (AsA) and reduced glutathione (GSH). Phenylalanine ammonia lyase (PAL) significantly increased in inoculated leaves of both cultivars, resulting in higher levels of total phenolic compounds and flavonoids. The pathogenesis‐related proteins chitinase (CHT) and β‐1, 3‐glucanase (GLU) significantly increased following inoculation with higher activity in the resistant cultivar. These findings show that resistance of muskmelon plants against C. lagenarium is associated with the rapid accumulation of H₂O₂, resulting in altered cellular redox status, accumulation of pathogenesis‐related proteins, activation of phenylpropanoid pathway to accumulation of phenolic compounds and flavonoids.     

The Response of Lettuce Cultivars against Two Lettuce mosaic virus isolates,One Able to Systemically Infect the Resistant Cultivar Salinas 88

The response of seven lettuce cultivars to two geographically different Lettuce mosaic virus (LMV) isolates (LMV‐A, LMV‐T) was statistically evaluated based on infection rate, virus accumulation and symptom severity in different time trials. LMV‐A is characterized by the ability to systemically infect cv. Salinas 88 (mo1²‐carrying resistant cultivar), and inducing mild mosaic symptoms. Among lettuce cultivars, Varamin (a native cultivar) similar to cv. Salinas showed the most susceptibility to both LMV isolates, whereas another native cultivar, Varesh, was tolerant to the virus with minimal viral accumulation and symptom scores, significantly different from other cultivars at P < 0.05. LMV‐A systemically infects all susceptible lettuce cultivars more rapidly and at a higher rate than LMV‐T. This isolate accumulated in lettuce cultivars at a significantly higher level, determined by semiquantitative ELISA and induced more severe symptoms than LMV‐T isolate at 21 dpi. This is the first evidence for a LMV isolate with ability to systemically infect mo1²‐carrying resistant cultivar of lettuce from Iran. In this study, accumulation level of LMV showed statistically meaningful positive correlation with symptom severity on lettuce plants. Based on the results, three evaluated parameters differed considerably by lettuce cultivar and virus isolate.     

Components of Resistance to Early Blight in Four Potato Cultivars: Effect of Leaf Position

Components of early blight resistance were quantified in leaves of different ages in four potato cultivars. The components of resistance: incubation period (IP), lesion number (LN), early blight severity, lesion expansion rate (LER), latent period (LP) and spore production by lesion area (SPLA), were evaluated separately in the lower, middle and upper leaves of four potato cultivars. Plants of cultivar Aracy (resistant), Delta (moderately resistant), Desirée (susceptible) and Bintje (susceptible) were inoculated with an Alternaria solani isolate at the beginning of the flowering stage. Disease severity varied in different plant parts. In all cultivars, regardless of resistance, the smallest values of LN, and severity were recorded on the upper leaves, suggesting that young tissues are less susceptible. In cultivar Aracy, the IP was long, with small values of LN and LER and consequently, low values of early blight severity in all leaf positions were recorded. Although IP was long in cultivar Aracy, no differences between the moderately resistant cultivar Delta and the susceptible cultivars Bintje and Desirée could be detected for this component. The IP was only influenced by leaf position in cultivar Aracy. Clear differences in resistance levels among cultivars could be detected regarding LN, severity and LER. However, neither LP nor SPLA were associated with resistance level of cultivars or with leaf position. Analyses according to plant part suggest that evaluations on leaves of the middle third part are most suitable for screening for early blight resistance in potato.     

Engineering fire blight resistance into the apple cultivar ‘Gala’ using the FB_MR5 CC‐NBS‐LRR resistance gene of Malus × robusta 5

The fire blight susceptible apple cultivar Malus × domestica Borkh. cv. ‘Gala’ was transformed with the candidate fire blight resistance gene FB_MR5 originating from the crab apple accession Malus × robusta 5 (Mr5). A total of five different transgenic lines were obtained. All transgenic lines were shown to be stably transformed and originate from different transgenic events. The transgenic lines express the FB_MR5 either driven by the constitutive CaMV 35S promoter and the ocs terminator or by its native promoter and terminator sequences. Phenotyping experiments were performed with Mr5‐virulent and Mr5‐avirulent strains of Erwinia amylovora, the causal agent of fire blight. Significantly less disease symptoms were detected on transgenic lines after inoculation with two different Mr5‐avirulent E. amylovora strains, while significantly more shoot necrosis was observed after inoculation with the Mr5‐virulent mutant strain ZYRKD3_1. The results of these experiments demonstrated the ability of a single gene isolated from the native gene pool of apple to protect a susceptible cultivar from fire blight. Furthermore, this gene is confirmed to be the resistance determinant of Mr5 as the transformed lines undergo the same gene‐for‐gene interaction in the host–pathogen relationship Mr5–E. amylovora.     

Role of Pseudomonas fluorescens and INA against Black Rot of Cabbage

Cabbage (Brassica oleracea var. capitata) is an important vegetable crop among crucifers. It is affected by a bacterial disease known as black rot. Black rot is caused by Xanthomonas campestris pv. campestris a disease of worldwide importance. The present study highlights the effect of biotic inducer—Pseudomonas fluorescens—and an abiotic inducer—2,6‐dichloro‐isonicotinic acid—in combating black rot, followed by their effect on the seed treatment and disease incidence, role of antioxidant enzymes followed by validation of the defence‐related genes by quantitative real‐time PCR. The resistant (Pusa mukta) and the highly susceptible (NBH boss) cabbage cultivars were analysed for defence‐related enzymes such as peroxidase and superoxide dismutase. An increase in total peroxidase and superoxide dismutase activity was observed upon inoculation with X. campestris pv. campestris. The activity was greater in resistant cultivar when compared to susceptible ones. Both enzyme activity assays and qPCR analyses for the expression of the defence genes in susceptible and resistant cultivars demonstrated that the peroxidase gene was up‐regulated in resistant cultivar compared to susceptible cultivar. The present study proved that P. fluorescens‐induced resistance against X. campestris pv. campestris in cabbage seedlings is more efficient as compared to the use of INA—abiotic inducer.     

甘蔗鞭黑粉菌侵染甘蔗的早期可视化观察

【目的】甘蔗鞭黑粉菌(Sporisorium scitamineum)引起的甘蔗黑穗病是我国甘蔗生产重要的病害。示踪甘蔗鞭黑粉菌侵染甘蔗的过程将有助于揭示其致病性和甘蔗抗黑穗病机制,为抗病品种的选育以及黑穗病的防治奠定基础。【方法】利用农杆菌介导的遗传转化技术对甘蔗鞭黑粉菌进行黄色荧光标记,对转化子进行配合及致病力检测,将标记菌株接种甘蔗感病品种ROC22及抗病品种中蔗1号、中蔗6号和中蔗9号并进行早期可视化观察。【结果】组成型表达的eYFP不影响标记菌株的配合及致病能力,而且黄色荧光性状能通过冬孢子稳定遗传。激光共聚焦显微观察表明,注射接种病原菌第5天,在感病品种ROC22的生长点已可见荧光菌丝及少量聚集状菌丝体,在抗病品种中蔗1号、中蔗6号和中蔗9号中可见少量单一丝状菌丝,无聚集状菌丝体。接种后35 d,在ROC22中可见大量聚集状菌丝体,但在中蔗品种中的聚集状菌丝体明显较少,而以中蔗1号最少。【结论】成功构建了甘蔗鞭黑粉菌侵染甘蔗的荧光示踪系统,并发现中蔗系列品种存在抑制甘蔗鞭黑粉菌菌丝体在细胞间扩展的机制。     

Biocontrol of Ascochyta blight by Azospirillum sp. depending on the degree of resistance of chickpea genotypes

Throughout arable land that was devoted to chickpea (Cicer arietinum L. (Family: Leguminosae) production, Ascochyta blight (Ascochyta rabiei (Pass.) L. (Order: Sphaeriales; Family: Mycosphaerellaceae) is a widespread disease that would lead to significant loss of chickpea yield. This study's purpose was to explain the responses of a resistant chickpea cultivar (ICC 12004) and a susceptible cultivar (Bivanij) in terms of disease resistance, disease symptoms appearance and expression pattern of two defence‐related genes (DEF0442 and Snakin2) after the Azospirillum brasilense seeds inoculation. In this research, the Snakin2 gene expression was affected by Azospirillum inoculation. The gene expression has been enhanced in plants inoculated with Azospirillum in both cultivars in comparison with non‐inoculated plants, but this change in ICC 12004 and Bivanij were significant and non‐significant, respectively. Although, Azospirillum would up regulate the DEF0422 gene expression in ICC 12004, but it would down regulate the expression of this gene in Bivanij. A. brasilense inoculation decreased the A. rabiei disease severity, regardless of the chickpea cultivar. Bivanij still could be classified as susceptible, even if treated with A. brasilense.     

Molecular cloning of phenylalanine ammonia-lyase cDNA and classification of varieties and cultivars of tea plants (Camellia sinensis) using the tea PAL cDNA probe

Tea (Camellia sinensis) phenylalanine ammonia-lyase (PAL) cDNA was cloned using labelled rice PAL cDNA as a probe. The PAL genes of the tea plant were investigated by restriction fragment length polymorphism (RFLP) analysis using tea PAL cDNA. PAL genetic variation in tea plants was much larger than predicted due to the presence of various hybridized fragments in the Assam hybrids, which are hybrids between C. sinensis var assamica and var sinensis. On the other hand, hybridized band patterns of Japanese green tea cultivars belonging to var sinensis could be divided into five groups. Furthermore, a short-length PAL probe, about 280 bp including the 3 untranslated sequence, detected 3 DNA fragments of different lengths, which were named A, B and D. An experiment tracing the PAL gene heredity showed that A, B and D fragments were inherited according to the Mendelian monogenic ratio. Therefore, PAL genes identifiable by A, B and D fragments are multiple alleles, and the PAL gene is present as a single gene in the tea haploid genome. It was also clear that five groups of Japanese green tea cultivars were characterized by the composition of these PAL fragments. From RFLP analysis using tea PAL cDNA, we succeeded in distinguishing Assam hybrids and Japanese green tea cultivars with high and low catechin content, respectively, and in grouping Japanese green tea at the cultivar level.     

Alternaria Brassicae Induced Changes in the Activity of Cell Wall Degrading Enzymes in Leaves of Brassica Juncea

After inoculation of Brassica juncea leaves with Alternaria brassicae, activities of the cell wall degrading enzymes, polygalacturonase (EC 3.2.1.15) and cellulase (EC 3.2.1.4) decreased in leaf blight resistant cultivar RC-781 and increased in the susceptible cultivar Varuna upto 3 d. In the leaves of both the cultivars 11 poly-peptides were observed in the absence of A. brassicae inoculation. After inoculation in the resistant cultivar RC-781 there was no change in the polypeptide pattern, while in the susceptible cultivar Varuna, four polypeptides (43.7 to 58.8 kDa) disappeared only at 3^rd day after inoculation. This revised version was published online in June 2006 with corrections to the Cover Date.     

Improving chickpea yield by incorporating resistance to ascochyta blight

Ascochyta blight [Ascochyta rabiei (Pass.) Lab.] is the most destructive disease of chickpea (Cicer arietinum L.), but it can be managed effectively by the use of resistant cultivars. Therefore, a breeding programme was initiated during 1977–78 at ICARDA, Syria, to breed blight-resistant, high-yielding chickpeas with other desirable agronomic traits. Crosses were made in main season at Tel Hadya, Syria, and the F₁s were grown in the off season at Terbol, Lebanon. The F₂, F₄ and F₅ generations were grown in a blight nursery in the main season where blight epidemic was artificially created. The plants and progenies were scored for blight resistance and other traits. The F₃ and F₆ generations were grown in the off season under normal day length to eliminate late-maturing plants. The pedigree method of breeding was followed initially, but was later replaced by the F₄-derived family method. The yield assessment began with F₇ lines, first at ICARDA sites and later internationally. A total of 1584 ascochyta blight-resistant chickpea lines were developed with a range of maturity, plant height, and seed size not previously available to growers in the blight-endemic areas in the Mediterranean region. These included 92 lines resistant to six races of the ascochyta pathogen, and 15 large-seeded and 28 early maturity lines. New cultivars produced 33% more seed yield than the original resistant sources. The yield of chickpea declined by 340 kg ha^-1, with an increase in blight severity by one class on a 1–9 scale, reaching zero yield with the 8 and 9 classes. Development of blight-resistant lines made the introduction of winter sowing possible in the Mediterranean region with the prospect of doubling chickpea production. Twenty three cultivars have been released so far in 11 countries.Joint contribution from ICARDA and ICRISAT. ICRISAT Journal Article no. JA 1886.