Microsatellite Markers Associated with Spot Blotch Resistance in Spring Wheat

Spot blotch, caused by Cochliobolus sativus, is a serious wheat (Triticum aestivum L.) disease in the warm areas of South Asia. Breeding for resistance in the past 15 years has produced limited progress, and newly developed wheat cultivars suffer considerable yield reductions under spot blotch epidemics in the region. Resistance is often controlled by multiple genes with additive effects. Marker‐assisted selection, in combination with field selection, could accelerate the identification of progeny with multiple genes for resistance early in the breeding process. A study was conducted to determine microsatellite markers associated with resistance in the F₇ progeny from a cross between the spot blotch‐susceptible Sonalika and resistant G162 wheat genotypes. A parental survey using 171 simple sequence repeats (SSR) primer sets and spread over 21 chromosomes of wheat identified 52% polymorphic loci. However, only 15 polymorphic markers showed association with two bulks, one each of progeny with low and with high spot blotch severity. The detailed analysis indicated that progeny lines with low spot blotch severity could be separated from those with high severity using three SSR markers located on three wheat chromosomes. The findings may be useful in developing a marker‐assisted selection strategy for spot blotch resistance in wheat.     

Wheat Resistance to Spot Blotch Potentiated by Silicon

Spot blotch, caused by the fungus Bipolaris sorokiniana, is one of the most important diseases on wheat. The effects of silicon (Si) on this wheat disease were studied. Plants of wheat cultivars BR‐18 and BRS‐208 were grown in plastic pots containing Si‐deficient soil amended with either calcium silicate (+Si) or calcium carbonate (?Si). The content of Si in leaf tissue was significantly increased by 90.5% for the +Si treatment. There was no significant difference between Si treatments for calcium content, so variations in Si accounted for differences in the level of resistance to spot blotch. The incubation period was significantly increased by 40% for the +Si treatment. The area under spot blotch progress curve, number of lesions per cm² of leaf area, and real disease severity significantly decreased by 62, 36 and 43.5% in +Si treatment. There was no significant effect of Si on lesion size. The role played by total soluble phenolics in the increased resistance to spot blotch of plants from both cultivars supplied with Si was not clear. Plants from cultivar BR‐18 supplied with Si showed the highest values for concentration of lignin‐thioglycolic acid derivatives during the most advanced stages of fungus infection. Chitinase activity was high at the most advanced stages of fungus infection on leaves from both cultivars supplied with Si and may have had an effect on fungus growth based on the reduction of the components of resistance evaluated. Peroxidase activity was found to be high only at 96 h after inoculation of both cultivars supplied with Si. Polyphenoloxidase activity had no apparent effect on resistance regardless of Si treatments. Results revealed that supplying Si to wheat plants can increase resistance against spot blotch.     

Genetic Improvement and Crop Management Strategies to Minimize Yield Losses in Warm Non-traditional Wheat Growing Areas due to Spot Blotch Pathogen Cochliobolus sativus

Spot blotch caused by Cochliobolus sativus emerged as a major threat to wheat production in the warmer non-traditional wheat growing areas in the late 1980s. This foliar disease causes significant yield losses annually (15–20% on average in South Asia) endangering the livelihoods of millions of small farmers. Effective measures in the field are needed to mitigate the impact of spot blotch on food security in affected areas. This review summarizes the global knowledge on genetic improvement and crop management strategies to minimize yield losses based on latest field research. Recent studies have shown that spot blotch severity is highly influenced by stress factors affecting crop physiology which in turn affects host tolerance and resistance to the pathogen. Soil nutrient and water stress aggravate spot blotch-induced grain yield losses. Heat stress which is gradually increasing in Asia causes higher levels of disease damage. Genetic improvement is the cornerstone of a sustainable control of spot blotch in all affected regions. Resistance is essentially based on Chinese and South American sources and inter-specific crosses with broadly adapted semi-dwarf germplasm. A list of genotypes consistently reported in the last 10 years to harbor at least partial resistance to spot blotch, along with their inheritance of resistance, has been compiled to help breeding programmes. As the fungus is aggressive under conditions of high relative humidity and heat which in turn influences plant susceptibility, a synthesis of the different tools for scoring disease severity is given. Because resistance is incomplete, the ultimate goal is the accumulation of minor genes of resistance in adapted high yielding genotypes. This paper shows how the use of resistant varieties, timely seeding, adequate fertilization, crop rotation, and the judicious use of fungicides can be part of an integrated management strategy for controlling yield losses due to spot blotch.     

The Association Between Leaf Malondialdehyde and Lignin Content and Resistance to Spot Blotch in Wheat

Spot blotch (causative pathogen Bipolaris sorokiniana (Sacc.) Shoem) is a common disease of wheat in the Eastern Gangetic Plains region of India. The association of leaf malondialdehyde and lignin contents with the severity of spot blotch disease was studied using a correlation analysis based on a population of recombinant inbred lines bred from the cross cvs. Yangmai 6 (resistant) × Sonalika (susceptible). The material was field‐tested over two consecutive years and inoculated artificially with a highly virulent strain of the pathogen. Disease severity was assessed at three growth stages around and after anthesis. Leaf lignin content tended to be higher in the more resistant RILs, while the opposite was the case for leaf malondialdehyde content. Lesion size showed a positive correlation with disease severity and leaf malondialdehyde content, while disease severity and leaf lignin content were negatively correlated with one another, as were leaf malondialdehyde and leaf lignin content. Leaf malondialdehyde and/or leaf lignin content could be informative as markers for selection for higher levels of resistance against spot blotch in wheat.     

URP‐based DNA Fingerprinting of Bipolaris sorokiniana Isolates Causing Spot Blotch of Wheat

Spot blotch, caused by the pathogen Bipolaris sorokiniana is an important disease of wheat and is responsible for large economic losses world wide. In this study, molecular variability in B. sorokiniana isolates collected from different regions of India was investigated using URP‐PCR technique. All the 40 isolates used in the study were pathogenic when tested on susceptible host, Agra local, although they varied in pathogenicity. Isolate BS‐49 was least virulent showing 4.5 infection index while BS‐75 was the most virulent with 63.4 infection index. The universal rice primers (URPs’) are primers which have been derived from DNA repeat sequences in the rice genome. Out of the 12 URP markers used in the study, 10 markers were effective in producing polymorphic fingerprint patterns from DNA of B. sorokiniana isolates. The analysis of entire fingerprint profile using unweighted pair group method with arithmetic averages (UPGMA) differentiated B. sorokiniana isolates obtained from different geographic regions. One isolate BS‐53 from northern hill zone was different from rest of the isolates showing less than 50% similarity. Broadly, three major clusters were obtained using UPGMA method. One cluster consisted of isolates from North western plain zone; second cluster having isolates from North eastern plain zone and third cluster consisted of isolates from Peninsular zone showing more than 75% genetic similarity among them. One of the markers, URP‐2F (5′GTGTGCGATCAGTTGCTGGG3′) amplified three monomorphic bands of 0.60, 0.80 and 0.90 kb size which could be used as specific markers for identification of B. sorokiniana. Further, based on URP‐PCR analysis, the grouping of the isolates according to the geographic origin was possible. This analysis also provided important information on the degree of genetic variability and relationship between the isolates of B. sorokiniana.     

Effect of Helminthosporic Root Rot on the Lipid Content in Wheat Seedlings

The concentration of dry matter and the content of esterified fatty acids in total lipids of roots and etiolated shoots of 3- to 10-day-old seedlings of wheat (Triticum aestivum L.) infected with Bipolaris sorokiniana (Sacc.) Schoemaker, the agent of helminthosporic root rot, were determined in the course of germination. At the onset of germination, fungal infection caused a considerable increase in the dry matter concentration in both roots and shoots due to the enhanced mobilization of seed reserves. However, after the 7th day of germination, dry matter concentration fell below the level of noninfected control seedlings as a result of infection. The content of total lipids rose immediately after infection and always exceeded the control index up to the end of germination, in spite of a continuous decrease in this index in both control and infected seedling. It is concluded that an increase in the content of cellular lipids is a characteristic response of both shoots and roots to the root rot infection of wheat seedlings.     

Virulence Spectrum of Mycosphaerella graminicola Isolates on Wheat Genotypes Carrying Known Resistance Genes to Septoria tritici Blotch

The virulence spectrum of 23 monopycnidiospore isolates of Mycosphaerella graminicola was determined using wheat genotypes that carried different resistance genes (Stb1–Stb8 and Stb15). Disease severity was measured as the percentage of necrotic leaf area. The isolates used in the experiments were of diverse origin: eight from Poland, seven from Germany, and eight from other countries around the world. Analysis of variance revealed significant differences in the virulence of the isolates. Using multiple regression and Cook’s D statistic, 26 significant cultivar × isolate interactions were detected. The Israeli isolate IPO86036 showed the widest spectrum of specific reactions. It expressed specific virulence on at least four cultivars and specific avirulence on at least three. The other isolates showed specific interactions with 1–6 different cultivars. Despite the limited number of isolates that were tested, we recommend that a number of resistant lines, namely cultivars Veranopolis (Stb2), Cs/Synthetic 7D (Stb5), Arina (Stb15, Stb6 and partial resistance), and Liwilla (unknown resistance factors), could be incorporated into central European wheat breeding programmes that are aimed at developing resistance against septoria tritici blotch. In contrast, resistance gene Stb7, which is carried by cultivar Estanzuela Federal, was ineffective against most of the isolates that were used. These results on the virulence spectrum of M. graminicola isolates provide valuable information for effective wheat breeding programmes to develop resistance to the pathogen.     

Cytological Study of Wheat Spike Infection by Bipolaris sorokiniana

The infection of wheat spikelets by Bipolaris sorokiniana , the causal agent of black point on grains and grain shrivelling, was examined by light and electron microscopy. Conidia of the pathogen germinated 6–12 h after inoculation on the surfaces of the different spike tissues. Extracellular sheaths were observed on germ tubes and appressoria attached to the surfaces of lemma, palea and seeds, but were only scarcely detected on the surface of conidia. Appressoria, frequently found over grooves, formed penetration hyphae invading the epidermal cell walls. Infection process was similar on the surface of the lemma, palea and glume. Growth of the fungus in the epidermal and parenchyma cells was found predominantly in the cell walls, and hyphae also extended intercellularly and intracellularly. Infection of seeds appeared to occur via two ways: (i) direct infection of the outer layers of the cell walls of the pericarp and (ii) through entering the stigma into the pericarp cells. Secretion of host cell wall hydrolytic enzymes at the apex of the penetrating hyphae may facilitate the spread of the fungus. In addition, toxins secreted by the fungus might explain the rapid death of host cells in contact with or distant to fungal cells. A host response to fungal infection involved the development of appositions between cell wall and plasma membrane in cells adjacent to fungal cells. Fungal hyphae were sometimes also surrounded by electron dense material.     

The Effect of Helminthosporic Root Rot on the Composition of Acyl Lipids of Wheat Seedlings

The changes of the molecular species composition of esterified fatty acids (FAs) of total and nonextractable lipids were determined in roots and etiolated shoots of 3- to 10-day-old wheat (Triticum aestivumL.) seedlings infected with the fungus Bipolaris sorokiniana, the agent of helminthosporic root rot. A novel technique of assessing the extent of the infection-induced deviation of FA composition, mol %, from the control value was developed. It consists in the quantitative determination of both the deviations in this composition and the extent of contribution of separate FA species to the deviations observed. The application of this technique has shown that, for the total lipids, the maximum of such a deviation, in accordance with the membrane theory of stress, directly coincided in time with the onset of a decrease in the dry matter content in both roots and shoots. In each of these, the deviation was primarily caused by the change in the content of those FA species that usually dominate in a specific group of membrane lipids prevailing in a given organ, viz., plastid glycolipids in shoots and extraplastidal membrane phospholipids in roots. In both cases, C₂₀–C₂₂FAs significantly contributed to the deviations observed. This fact seems to reflect an enhanced formation of epicuticular waxes rich in these FAs on the shoot and root surfaces as an adaptive response of plants to fungal infection. Nonextractable (annular) membrane lipids, because of their vital importance for the survival of plant cells, differed from the total lipids with a far greater stability of their quantitative FA composition under conditions of infection-induced metabolic disturbances.     

Wheat Resistance to Bacterial Leaf Streak Mediated by Silicon

Leaf streak, caused by Xanthomonas translucens pv. undulosa, is the major bacterial disease of wheat in Brazil and other countries worldwide. This study aimed to evaluate the effect of silicon (Si) on disease development and the biochemical mechanisms possibly involved in resistance potentialized by this element. Plants of cv. BR‐18, susceptible to leaf streak, were grown in plastic pots containing Si‐deficient soil amended with either calcium silicate (+Si) or calcium carbonate (?Si). The content of Si increased (P ≤ 0.05) by 96.5% for the +Si when compared with ?Si treatment. There was no difference (P ≥ 0.05) between Si treatments for calcium content on leaf tissue, so variations in Si accounted for differences in the level of resistance to leaf streak. There was no difference (P ≥ 0.05) between Si treatments for incubation period, latent period, necrotic leaf area, and severity estimated by the software quant . However, chlorotic leaf area was reduced (P ≤ 0.05) by 50.2% for the +Si when compared with ?Si treatment. There was no difference (P ≥ 0.05) between Si treatments for the bacteria population on leaf tissue; however, the values seemed to be somewhat lower in the +Si treatment from 4 to 8 days after inoculation (d.a.i.) on leaves from plants supplied with Si. There was no difference (P ≥ 0.05) between Si treatments for electrolyte leakage. The concentration of total soluble phenolics and lignin‐thioglycolic acid (LTGA) derivatives did not show any apparent signs of increase during the course of infection and seemed to be slightly higher on plants not supplied with Si at the most advanced stages of bacterial infection. Chitinase activity was high at the most advanced stages of bacterial infection on leaves from +Si treatment and probably affected bacterial growth on leaf tissue. Peroxidase activity following bacterial infection was not increased by Si, but can be linked with the highest concentration of LTGA derivatives at 12 d.a.i. of plants supplied with Si. Polyphenoloxidase activity did not affect wheat resistance to leaf streak regarding of the Si treatments. The results clearly suggest that supplying Si to wheat plants can increase resistance to leaf streak possibly through an increase in tissue lignification and the participation of chitinases and peroxidases.     

Rice Resistance to Brown Spot Mediated by Nitrogen and Potassium

This study was undertaken to investigate the effects of both nitrogen (N) and potassium (K) rates on rice resistance to brown spot, caused by the fungus Bipolaris oryzae. Rice plants (cultivar ‘Metica 1’) were grown in soil corrected with 0, 25, 50, 75 and 100 mg of N / kg (as NH₄NO₃) of soil as well as with 25, 50, 75, 125 and 150 mg of K / kg (as KCl) of soil. Thirty‐three‐day‐old plants were inoculated with a suspension of Bipolaris oryzae conidia and the incubation period (IP), number of lesions (NL) per cm² of leaf area and disease severity was evaluated. Disease severity was scored at 24, 48, 72, 96, 120 and 144 h after inoculation and data were used to obtain the area under brown spot progress curve (AUBSPC). Soil plant analysis development (SPAD) index, plant dry weight and concentration of N and K in leaf tissues were also determined for both non‐inoculated (NI) and inoculated (IN) plants. Concentration of N in leaf tissue increased as the N rates in the soil increased. Concentration of K in leaf tissue increased sharply as the K rates in the soil increased for both NI and IN plants. Concentration of K in leaf tissue was not affected by N rates. The IP increased as the N rates increased, but was somewhat less impacted by increasing K rates. The NL decreased as the N rates increased. The NL dramatically declined at the highest K rates. The AUBSPC dramatically declined as the N and K rates in the soil increased. SPAD index values increased as the N and K rates in the soil increased for both NI and IN plants. Plant dry weight increased as the N and K rates in the soil increased for both NI and IN plants. Results from this study suggest that combining high N and K rates may contribute to reducing the intensity of brown spot in rice while improving plant development.     

The Effect of Local Cooling of Cucumber and Wheat Seedlings on Various Kinds of Stress Resistance of Their Leaves and Roots

Shoots and roots of wheat (Triticum aestivum L., cold-resistant species) and cucumber (Cucumis sativus L., cold-sensitive species) were chilled at 2°C or 10°C, respectively, for 7 h. The changes in cold, heat, and salt resistance in treated leaf and root cells were recorded. Local cooling of the leaf resulted in an increase of its cold and salt tolerance, but its heat tolerance remained unchanged. At the same time, cold tolerance of the root slightly increased as a result of local cooling, but its heat and salt tolerance decreased. Cooling of the shoot did not affect the cold and heat tolerance of root cells but caused a decrease in their salt tolerance. Finally, in the leaf maintained at a normal temperature, there was an increase in all kinds of stress resistance as a result of root cooling. We discuss the possibility of an unspecific change in stress resistance caused by metabolic shifts. These shifts are induced by a signal, which is transmitted inside the plant into plant organs located at a considerable distance from the chilled ones.     

小麦PAL基因的克隆及赤霉菌诱导下的表达分析

利用苯丙氨酸解氨酶(PAL,phenylalanine ammonia-lyase)基因保守区域从小麦抗赤霉病材料苏麦3号中克隆获得4个PAL基因,分别命名为Ta PAL1、Ta PAL2、Ta PAL3、Ta PAL4。4个基因的开放阅读框(ORF,open reading frame)长度分别为2142 bp、2016 bp、2118 bp和2139 bp,分别编码714个、672个、706个和713个氨基酸。基因序列比对发现其相似性达到88.35%,所编码的氨基酸相似性为91.92%,氨基酸序列分析表明4个基因都包含HAL-PAL结构域及PAL结构域。通过接种禾谷镰刀菌,利用荧光定量PCR对PAL基因进行表达分析发现,4个PAL基因全部为上调表达,其中Ta PAL2、Ta PAL3和Ta PAL4最为明显。PAL基因的上调表达,说明PAL基因在小麦抵抗赤霉病菌侵染的机制中可能起着重要作用。     

Minor Differences in Anther Extrusion Affect Resistance to Fusarium Head Blight in Wheat

Fusarium head blight (FHB) remains a serious problem due to yield loss and mycotoxin accumulation in wheat production worldwide. We previously reported that the closed‐flowering (no anther extrusion) characteristic was effective for increasing resistance to FHB infection. In this study, we investigated the relationships between the degree of anther extrusion (AE) and FHB damage using double haploid lines (DHLs), derived from F₁ plants from crosses between closed‐flowering and opened‐flowering varieties. These DHLs exhibited various degrees of AE, and the degree of AE was significantly different among DHLs, regardless of the year and environment (pot‐ or field‐grown). FHB severity was the lowest in closed‐flowering DHLs, and DHLs with partially extruded anthers showed significantly higher FHB symptoms than those with closed‐flowering phenotypes. In general, DHLs with partially extruded anthers also had relatively severe FHB symptoms compared with those exhibiting full anther extrusion. FHB severity was significantly correlated with Fusarium‐damaged kernels and deoxynivalenol concentration. The results of this study showed that partially extruded anthers were considered to be a source of FHB infection. The closed‐flowering phenotype improved resistance to FHB infection. Meanwhile, phenotypes with rapid anther extrusion and ejection also could contribute to the avoidance of FHB infection.     

HRGP在小麦抗寒锻炼过程中的变化及其与抗寒性的关系

强抗寒小麦品种(R-025、中品94-19、品83-1、品83-2、品83-3、米罗诺夫808)经抗寒锻炼后,其幼苗体内的游离脯氨酸、细胞壁结合的羟脯氨酸和糖蛋白含量发生了明显的变化.游离脯氨酸含量比未经抗寒锻炼处理时增加5～32倍,细胞壁结合的羟脯氨酸含量比对照增加1.77～2.17倍,糖蛋白含量比对照增加4.68～9.72倍,而不抗寒小麦品种(中国春、冬103)增加量较小.脯氨酸积累进程各个品种间差异比较大,品83-1、品83-2积累较快,抗寒锻炼第21d时达到最高峰,而R-025在第56d达到最高峰.脯氨酸含量与小麦品种抗寒性相关不显著(相关系数为0.3462),而羟脯氨酸含量、糖蛋白含量与小麦品种抗寒性相关显著,相关系数分别为0.6491和0.7039.从小麦细胞壁纯化得到了2种伸展蛋白Extensm1和Ex-tensin2,其含量都和小麦品种抗寒性呈正相关.Extensin1是分子量为28kD、羟脯氨酸为主要成份(32mo1%)的富含羟脯氨酸糖蛋白.     

普通小麦99-2439中的白粉病抗性遗传

普通冬小麦品系99-2439在郑州连续4年对田间白粉菌(Blumeria graminis sp.tritici)表现高抗,但其抗性基因来源不清.通过染色体C-分带和IRS染色体特异性SCAR标记鉴定,表明它是一个小麦-黑麦(Triticum aestivum-Secale cereale)lBL/1RS异易位系.通过对中国春×99-2439杂交F2代分离群体抗性鉴定和1RS染色体臂检测结果分析,证明该抗病基因不在1RS染色体臂上.用单孢小麦白粉菌分离株对其抗性遗传进行研究,结果表明,99-2439的白粉病抗性由一对小种专化、隐性抗病基因控制.由于携带Pm5a的Hope/8Cc对中国的21个小麦白粉菌分离菌株均高度感病,而99-2439高抗混和白粉菌和5个单孢分离菌株,所以,99-2439所携带的抗白粉病基因不同于Pm5a.     

Dry Matter Partitioning between Plant Organs in Wheat Cultivars Differing in Productivity and Drought Resistance

In microplot field experiments conducted over the course of 12 years, the accumulation of dry matter was recorded in the leaves, stems, and ears of the primary shoots of plants of four cultivars of spring wheat (Triticum aestivumL.) differing in productivity and drought resistance. The contribution of organs to the accumulation of dry matter by the shoot from emergence to anthesis was calculated, and relative changes in the weight of the ear after anthesis were assessed. In all the cultivars, the contribution of the leaves was the same; however, the share of the ear was greater in short-stem cultivars of the intensive type in which the leaves were more active during the time of grain filling. Furthermore, this load increased, and the relative increment in the ear weight after flowering decreased in the years of drought, because the contribution of the leaves did not depend on water supply during the growing season. During these years, the contribution of the ear increased in the plants of all the cultivars. The authors conclude that, in arid regions, in addition to drought resistance, the cultivar should display such a ratio between the ear weight and the weight of leaves that would still ensure satisfactory grain filling