Reaction of sugar beet S1 lines and cultivars to different isolates of Macrophomina phaseolina and Rhizoctonia solani AG-2-2IIIB

Interactions of 17 sugar beet lines and cultivars with four isolates of Macrophomina phaseolina (the causal agent of charcoal rot) and one isolate of Rhizoctonia solani (the causal agent of crown and root rot) were studied in separate experiments under greenhouse conditions. The isolates of Macrophomina were taken from their host plants, sugar beet (two isolates), soybean and sesame. In the first experiment, the colonized toothpick was used as inoculum. In the second experiment, six-month-old sugar beet plants were inoculated with barley seeds colonized with M. phaseolina. For the inoculation of sugar beet lines with R. solani, the colonized corn seeds were used. Root symptoms were recorded four weeks after inoculation, by estimating the proportion of the root surface infected by the pathogens, using a 1–9 standard scale. Our results showed a significant difference among lines and cultivars in their resistance to these two pathogens. Line B8618 was found to be considerably resistant to the isolates of the both pathogens. The inoculation methods of Macrophomina isolates had no significant effect on the results. The interaction between isolate and cultivar was not also significant in Macrophomina-resistant lines. Therefore, it appears that the response of sugar beet lines to the tested fungal isolates was not differential. These resistant lines showed a high resistance to all the tested M. phaseolina isolates. Our results revealed that the Macrophomina-resistant lines also showed resistance to R. solani. Furthermore, the sugar beet drought tolerant lines (M293, M362 and M345) were susceptible to the tested M. phaseolina and R. solani isolates.     

Molecular markers for improving control of soil-borne pathogen <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> in sugar beet

Fusarium spp. cause severe damage in many agricultural crops, including sugar beet, with Fusarium oxysporum historically being considered as the most damaging of all species. Sugar beet needs to be protected from this class of soil-borne pathogens in order to ensure an optimal sugar yield in the field. Genetic control of the disease is crucial in managing these pathogens. Identification of single nucleotide polymorphism (SNP) markers linked to resistance can be a powerful tool for the introgression of valuable genes needed to develop Fusarium-resistant varieties. A candidate gene approach was carried out to identify SNP markers linked to putative Fusarium resistance sources in sugar beet. Five resistant analogue genes (RGAs) were screened by means of high resolution melting (HRM) analysis in a set of sugar beet lines, considered as resistant and susceptible to Fusarium oxysporum. HRM polymorphisms were observed in 80% of amplicons. Two HRM polymorphisms were significantly associated with Fusarium resistance (P < 0.05). The amplicons that showed association were sequenced and two SNPs were identified. The association was further validated on 96 susceptible and 96 resistant plants using competitive allele-specific PCR (KASPar) technology. The selected SNPs could be used for marker-assisted breeding of Fusarium resistance in sugar beet.     

不同氮素水平对甜菜氮代谢酶和可溶性蛋白含量的影响

为了筛选合适的氮素施用量,研究氮素营养和氮素同化关键酶的关系,以尿素为氮源,采用HI003为试材,研究了不同的施氮水平对甜菜叶片、块根中可溶性蛋白含量、硝酸还原酶和谷氨酰胺合成酶活性的影响。结果表明：叶片中,硝酸还原酶的活性随着施氮水平的提高而增加,但是超过 200 kg/hm²酶活性下降;块根中硝酸还原酶活性在0~120 kg/hm²随着施氮量增加而上升,超过 160 kg/hm²酶活力下降。叶片、块根中,整个生育期施氮肥处理增加了谷氨酰胺合成酶活性。试验中发现施氮量和生育期早期的叶片中硝酸还原酶和谷氨酰胺合成酶活性呈显著正相关。施氮量和甜菜叶片可溶性蛋白含量在7月15日和9月15日呈显著正相关;在生育期内,氮素水平和块根中的可溶性蛋白含量呈极显著正相关。     

Expression in vitro of resistance to Heterodera schachtii in hairy roots of an alien monotelosomic addition plant of Beta vulgaris,transformed by Agrobacterium rhizogenes

Summary Hairy roots, induced by Agrobacterium rhizogenes, were obtained of nematode susceptible beet plants (Beta vulgaris) and of the nematode resistant alien monotelosomic addition AN5, carrying a telosome from B. patellaris. The additional telosome was found to be stably present in vitro in the roots of AN5. The hairy root cultures were inoculated with larvae of the beet cyst nematode Heterodera schachtii. On the root culture of AN5 significantly less cysts developed than on the other root cultures. These results indicate that the resistance to the beet cyst nematode is expressed in the roots after transformation and can be monitored under in vitro conditions.     

Wound response in orange as a resistance mechanism against Penicillium digitatum (pathogen) and P. expansum (non-host pathogen)

Penicillium digitatum is the most devastating postharvest pathogen of citrus. In addition, Penicillium expansum is the main pathogen of pome fruit, although recent studies have demonstrated its ability to infect oranges under some conditions. In this study, we evaluated wound response in ‘Valencia’ oranges harvested at three different maturity stages and the effect of wound response on the establishment of both pathogens when fruit were stored at two different temperatures (20 and 4 °C). The effect of wounding and pathogen inoculation on lignin content, was also quantified. Lastly, the expression of several phenylpropanoid pathway-related genes was also analyzed by semi-quantitative RT-PCR. Results indicated that, in general, P. digitatum exhibited lower decay incidence and severity as time between wounding and inoculation increased. Decay incidence and severity were higher in fruit from the over-mature harvest than in fruit from immature and commercial harvests. P. expansum was able to infect fruit at 20 °C but lesions were small compared to lesion size of fruit stored at 4 °C. Lignin content in wounded fruit (control) and in samples wounded and inoculated with P. expansum was highest in fruit from the immature harvest at 7 d post-wounding and inoculation. Wounded fruit had higher expression of pal1, comt1 and pox1 genes at 48 h than at 24 h. However, samples inoculated with P. digitatum showed lower expression at 48 h than at 24 h. Our results indicated that maturity and storage temperature play an important role in orange wound response.     

Evaluation of Verticillium wilt resistance in commercial cultivars and advanced breeding lines of cotton

Verticillium wilt (VW), caused by Verticillium dahliae Kleb, is one of the most destructive diseases in cotton (Gossypium spp.). The most efficient and cost-effective method of controlling the disease is the use of resistant cotton cultivars. Most commercial cultivars and elite breeding lines are developed under non-VW conditions and their responses to the disease are currently unknown. This study was conducted to evaluate current commercial cotton cultivars and advanced breeding lines for VW resistance. In 2011–2013, a total of 84 cultivars from major US seed companies, 52 advanced breeding lines from the US public breeding programs, and 87 introgression lines from a cross between Acala 1517-99 × Pima PHY 76 from the New Mexico Cotton Breeding Program, were evaluated for VW resistance in the greenhouse. Cotton cultivars and breeding lines were evaluated in ten separate replicated tests by inoculation with a defoliating-type isolate of V. dahliae. While leaf severity rating and percentages of infected plants, infected leaves and defoliated leaves were found to be significantly and positively correlated with one another, leaf severity rating and percentage of infected leaves were best choices because of their relatively low coefficients of variation and higher resolutions to differentiate resistant genotypes from susceptible ones. The heritabilities for the VW resistance traits ranged from 0.58 to 0.80 with an average of 0.67, indicating that variation in VW resistance is predominantly due to genetic factors. Of the 223 commercial cultivars and advanced lines, six Upland cultivars (FM 9160B2F, FM 9170 B2F, NG 4010 B2RF, Nitro 44 B2RF, DP 1219 B2RF, and ST 4288 B2F), five advanced lines (Ark 0403-3, MD 10-5, MD 25ne, NC11AZ01, and PD 0504), two introgression lines from Upland × Pima (NM11Q1157 and 08N1618), and four Pima cultivars (COBALT, DP 357, PHY 800, and PHY 830) had higher levels of resistance to VW. The resistance shown by most of these cultivars in the greenhouse was consistent with their performance in previous field tests. Based on the initial VW resistance, 19 highly or moderately resistant genotypes were chosen for re-evaluation and 30 genotypes were also assessed more than once for VW resistance in different tests, most of which had concordant performance. These cultivars and advanced lines should be useful resources to improve VW resistance in cotton breeding.     

Hybrid breeding for fall armyworm resistance: Combining ability and hybrid prediction

Fall armyworm (FAW, Spodoptera frugiperda) emerged as a major lepidopteran pest destroying maize in sub-Saharan Africa. A diallel mating design was used to generate 210 experimental hybrids from 21 lines. Experimental hybrids and four checks were evaluated in two locations. Commercial checks suffered higher foliar and ear damage compared to the top 15 hybrids. Mean squares associated with the genotypic variation were higher than genotype-by-environment interactions for foliar and ear damage traits. Heritabilities were moderate to high. Significant correlations were observed between grain yield (GY) with ear rot (−0.54) and ear damage (−0.45). Positive and significant GCA effects were observed for GY in seven parental lines, which were developed from multiple insect resistance breeding programmes. CKSBL10153 has the highest GCA value for GY and shows significant GCA effects for foliar and ear damage traits. These lines were identified as the ideal combiners for GY and FAW resistance and are therefore recommended for utilization as testers in the development of FAW-resistant three-way cross-hybrid maize with correlated response for increased GY. GCA and marker-based prediction correlations of GY were 0.79 and 0.96, respectively. Both GCA effects and marker-based models were effective in predicting hybrid performance for FAW resistance.     

New techniques for breeding maize (Zea mays) varieties with fall armyworm resistance and market-preferred traits for sub-Saharan Africa

Deploying maize varieties with fall armyworm (Spodoptera frugiperda [J.E. Smith]; FAW) resistance, desirable product profiles (PPs) and climate resilience is fundamental for food and economic security in sub-Saharan Africa (SSA). This study reviewed and identified challenges and opportunities for effective and accelerated breeding of demand-led maize hybrids with FAW resistance and adaptation to the diverse agro-ecologies of SSA. Lessons were drawn on improving breeding efficiency through adequate genetic variation delivered via prebreeding programmes, speed breeding and a reduced breeding stage plan. Appropriate PPs aligned with demand-led breeding approaches were highlighted as foundations for variety design and commercialization. Challenges to accelerated FAW resistance breeding in maize included inadequate funds and modern tools; poor adaptation of some exotic donor parental lines; lack of information on FAW resistance among local varieties; lack of integration of molecular markers associated with FAW resistance and agronomic traits into selection plans; and limited infrastructure for FAW rearing and germplasm screening. Integration of modern breeding tools and scientific innovations were recommended for accelerated development and release of FAW resistant and market-preferred maize varieties.     

甜菜M14品系BvM14-STPK基因的生物信息学分析及响应盐胁迫的表达分析

旨在进一步研究BvM14-STPK蛋白激酶对盐胁迫的应答反应,本研究以甜菜M14品系为材料,使用特异性引物通过聚合酶链式反应进行扩增,获得BvM14-STPK基因cDNA全长,并进行生物信息学分析,采用荧光定量PCR探究该基因响应盐胁迫的表达分析。生物信息学分析结果表明,BvM14-STPK基因cDNA全长1668 bp,包含1527 bp的最大ORF,编码508个氨基酸;BvM14-STPK蛋白激酶具有典型的蛋白激酶保守结构域。实时荧光定量PCR结果表明,BvM14-STPK基因在甜菜M14品系叶、根中均有广泛表达,叶中的表达量高于根中。该基因在200、400 mmol/L NaCl处理下,在叶片和根中呈现不同的表达状态,表明该基因可以应答盐胁迫,但在不同组织中应答盐胁迫的表达量存在差异。本项研究在挖掘甜菜M14品系优质基因和提高栽培甜菜抗逆性等方面具有重要指导意义,为进一步开展甜菜遗传改良工作奠定基础。     

Sources of resistance to <Emphasis Type="Italic">Phytophthora</Emphasis> root rot within the genus <Emphasis Type="Italic">Beta</Emphasis>

Phytophthora root rot caused by Phytophthora drechsleri Tucker is one of the most devastating sugar beet diseases in tropical areas. To identify genetic resources resistant to this disease, an aggressive isolate of P. drechsleri was selected. Then, a screening method was optimized based on the standard scoring scales of 1–9 (1: no symptoms, 9: complete plant death). Finally, 19 sugar beet lines, three cultivars, and 14 accessions of the wild species Beta vulgaris subsp. maritima, B. macrocarpa, B. procumbens, and B. webbiana were evaluated for resistance to the most aggressive isolate of P. drechsleri by using the optimized method (inoculum included 20 g of rice seed together with superficial wound creation). The isolates of P. drechsleri had significant variation in aggressiveness, and Kv10 was the most aggressive isolate on the susceptible variety Rasoul. The lines O.T.201-15, SP85303-0 (resistant check), and S2-24.P.107 had the lowest disease index with scores of 3.09, 3.13, and 3.27 respectively; they were categorized into the resistant group. The interaction between isolates and genotypes was not significant, which indicated the same response of each genotype to different isolates. Investigating the resistance of different generations of sugar beet revealed that progeny selection would be an effective method for increasing the resistance level of breeding materials to P. drechsleri. Among the wild species, the accession 9402 belonging to B. macrocarpa and the accession 7234 of B. vulgaris subsp. maritima had the lowest disease index (2.29 and 2.60, respectively) and were categorized into the resistant group.     

Inheritance of resistance to Xanthomonas campestris pv phaseoli in tetary bean (Phaseolus acutifolius)

Summary The F1, F2 and F3 from two crosses within Phaseolus acutifolius were exposed to Xanthomonas campestris pv phaseoli to analyse the inheritance of resistance. The resistant parent, PI 319.443, gave a hypersensitive reaction in leaves and pods with small necrotic lesions. Based on the resistance of F1, the segregation in F2 and the reaction of F3 plants and lines, it is concluded that resistance in leaves and pods is governed by one dominant gene. Comparisons are made with the resistance to X. campetris in P. vulgaris.     

Determining resistance to <Emphasis Type="Italic">Fusarium verticillioides</Emphasis> and fumonisin accumulation in African maize inbred lines resistant to <Emphasis Type="Italic">Aspergillus flavus</Emphasis> and aflatoxins

Fusarium verticillioides and Aspergillus flavus cause Fusarium ear rot (FER) and Aspergillus ear rot (AER) of maize, respectively. Both pathogens are of concern to producers as they reduce grain yield and affect quality. F. verticillioides and A. flavus also contaminate maize grain with the mycotoxins fumonisins and aflatoxins, respectively, which has been associated with mycotoxicosis in humans and animals. The occurrence of common resistance mechanisms to FER and AER has been reported. Hence, ten Kenyan inbred lines resistant to AER and aflatoxin accumulation were evaluated for resistance to FER, F. verticillioides colonisation and fumonisin accumulation; and compared to nine South African lines resistant to FER and fumonisin accumulation. Field trials were conducted at three localities in South Africa and two localities in Kenya. FER severity was determined by visual assessment, while F. verticillioides colonisation and fumonisin content were quantified by real-time PCR and liquid chromatography tandem mass spectrometry, respectively. Significant genotype x environment interactions was determined at each location (P ≤ 0.05). Kenyan inbred CML495 was most resistant to FER and F. verticillioides colonisation, and accumulated the lowest concentration of fumonisins across localities. It was, however, not significantly more resistant than Kenyan lines CML264 and CKL05015, and the South African line RO549 W, which also exhibited low FER severity (≤5%), fungal target DNA (≤0.025 ng μL^?1) and fumonisin levels (≤2.5 mg kg^?1). Inbred lines resistant to AER and aflatoxin accumulation appear to be promising sources of resistance to F. verticillioides and fumonisin contamination.     

Selection of Diploid Nematode-Resistant Sugar Beet from Monosomic Addition Lines

Monosomic sugar beet/wild beet addition lines (2n = 19) with full resistance against the beet cyst nematode have been characterized in different ways. Within the B. procumbens and B. webbiana addition lines three groups could be classified according to their isozymes pattern, growth habit, transmission rate, and resistance level. It is assumed that B. procumbens and B. webbiana each possess three different chromosomes which carry genes for nematode resistance. In the offspring of the addition lines diploid translocation types appear at very low frequencies, Isozyme pattern or growth type of the resistant plants were used for selecting diploid types in the offspring of monosomic addition lines. Effective selection could be made in progenies of susceptible sugar beets pollinated by addition lines because the pollen transmission of the alien chromosome is very low. Using these methods 7 nematode-resistant sugar beet lines could be selected. The transmission rates of the resistance gene ranged from 70.6% to 100%. Threw heterozygous progenies showed a 1:1 segregation indicating monogenic dominant inheritance of resistance. The level of resistance was as high as in the addition lines.     

条锈菌诱导的小麦MBF1转录辅激活因子基因的克隆及其特征分析

应用电子克隆和RT-PCR方法,从小麦叶片中分离出一个条锈菌诱导的编码MBF1基因的cDNA序列,暂被命名为TaMBF1a。TaMBF1a包含一个完整的429 bp的开放阅读框,编码142个氨基酸,具有MBF1保守结构域;小麦TaMBF1a氨基酸序列与水稻OsMBF1相似性达92%,与拟南芥AtMBF1a相似性达80%。TaMBF1a编码的蛋白可能是核蛋白,且该基因在小麦根、茎、叶组织中表达量基本一致。在小麦与条锈菌的亲和、非亲和互作中,TaMBF1a基因均受条锈菌诱导高水平表达,且非亲和组合表达量高于亲和组合。外源植物激素水杨酸、乙烯、脱落酸也可诱导该基因快速上调表达,表明TaMBF1a可能通过水杨酸、乙烯等信号途径参与小麦对条锈菌的防御反应。     

Overwintering of genetically modified sugar beet, Beta vulgaris L. subsp. vulgaris, as a source for dispersal of transgenic pollen

The potential impact of transgenic crops on community ecology will depend on the distribution and establishment of the new transgenic traits, on the sexual transfer of their new genes to the environment (Bartsch &; Pohl-Orf, 1996) and on the potential ecological impact of the transgenic trait. Flowering and pollen dispersal is important for outcrossing of the genetically engineered trait. For a biennial plant, like the cultivars of Beta vulgaris L., overwintering is normally necessary to become generative and to produce pollen and seeds (Abe et al., 1997), which usually does not happen with sugar beet as a field crop harvested in autumn (Longden 1989). The starting point for the project was a transgenic sugar beet, Beta vulgaris L. subsp. vulgaris (Lange et al., 1998), with rhizomania and herbicide ( Basta®, Liberty®) resistance. Cold tolerance is one of the most important factors for survival of sugar beet in Central- and North-Europe. Among other ways, spreading of transgenic traits into weed beet (Boudry et al., 1993) or wild beet can occur if genetically engineered – biennial – plants survive the winter, flower in spring and spread their pollen. Field experiments were performed with transgenic breeding lines and their hybrids, transgenic and non-transgenic hybrids with Swiss chard and three conventional beet cultivars to evaluate winter survival rates at seven different field sites. We could show that survival of sugar beet – transgenic as well as conventional ones – in Germany and at the Dutch border is possible. Survival rates were well correlated with temperature data and were unexpectedly high. Differences between sugar beet hybrids and breeding lines could be detected but not within different breeding lines or hybrids. There were no differences detectable between transgenic and non-transgenic plants. The data are crucial for the risk assessment of the release of transgenic sugar beet and are the basis for further experiments towards outcrossing and establishment.