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.     

Mapping of rhizoctonia root rot resistance genes in sugar beet using pathogen response‐related sequences as molecular markers

Rhizoctonia root and crown rot caused by the fungus Rhizoctonia solani is a serious disease of sugar beet. An F_2:3 population from a cross between a resistant and a susceptible parent has been tested for R. solani resistance and a genetic map has been constructed from the corresponding F₂ parents. The map encompasses 38 expressed sequence tags (ESTs) with high similarity to genes which are involved in resistance reactions of plants (R‐ESTs) and 25 bacterial artificial chromosomes (BACs) containing nucleotide binding site (NBS)‐motifs typical for disease resistance genes. Three quantitative trait loci (QTL) for R. solani resistance were found on chromosomes 4, 5 and 7 collectively explaining 71% of the total phenotypic variation. A number of R‐ESTs were mapped in close distance to the R. solani resistance QTL. In contrast, the NBS‐BACs mapped to chromosomes 1, 3, 7 and 9 with two major clusters of NBS‐BACs on chromosome 3. No linkage between NBS‐BACs and R. solani resistance QTL was found. The data are discussed with regard to using R‐ESTs and NBS markers for mapping quantitative disease resistances.     

Inheritance of Dry Root Rot (Rhizoctonia bataticola) Resistance in Chickpea (Cicer arietinum)

The inheritance of resistance to dry root rot of chickpea caused by Rhizoctonia bataticola was studied. Parental F₁ and F₂ populations of two resistant and two susceptible parents, along with 49 F₁ progenies of one of the resistant × susceptible crosses were rested for their reaction to dry root rot using the blotting-paper technique. All F, plants of the resistant × susceptible crosses were resistant; the F₂ generation fitted a 3 resistant: 1 susceptible ratio indicating monogenic inheritance, with resistance dominant over susceptibility. F³ family segregation data confirmed the results. No segregation occurred among the progeny of resistant × resistant and susceptible × susceptible crosses.     

黑龙江省大豆根腐病致病病原种类分布及抗病种质鉴定

为了明确黑龙江省大豆根腐病菌种类并筛选抗病种质资源,2008年以来,从黑龙江省大豆产区100个地点采集根腐病株,通过采用PDA培养基分离镰刀菌、立枯丝核菌、腐霉菌,选择培养基分离疫霉菌的方法。分离结果表明,各主要致病菌出现频率为：镰刀菌42.7%,腐霉菌25.2%,立枯丝核菌7%,疫霉菌25.2%。对1249份大豆材料在自然发病条件下鉴定的结果表明,病情指数15%以下的材料有471份,占鉴定材料的37.7%;对471份材料人工接种出现频率最高的镰刀菌的结果表明,病情指数15%以下的材料69份,占供试材料的14.6%。通过试验研究明确了大豆根腐病致病病原菌种类;明确了现行大豆品种和资源中有抗根腐病的材料。     

黑龙江省甜菜立枯病病区和品种抗病性分类研究

为了解决甜菜立枯病防治问题,2009—2016年,对黑龙江省11个地区157个甜菜品种,采取田间调查发病率的方法进行分类研究。将地区分为轻病区、中病区和重病区3类;将品种的抗性水平分为抗病、高抗、中抗、中感、高感、感病6个级别：甜菜立枯病发病率≤5%的地区为轻病区,5%<发病率≤15%的地区为中病区,发病率大于15%的地区为重病区。抗病品种发病率≤3%,3%<高抗品种发病率≤10%,10%<中抗品种发病率≤17%,17%<中感品种发病率≤24%,24%<高感品种发病率≤31%,感病品种发病率大于31%。依此标准划分出抗病品种24个,高抗品种61个,中抗品种49个,中感品种13个,高感品种3个,感病品种7个。依安、望奎、佳木斯、拜泉、宁安为轻病区,轻病区宜用中感、中抗、高抗、抗病品种;海伦、泰康为重病区,重病区宜用高抗和抗病品种;红兴隆、讷河、九三、呼兰为中病区,中病区宜用中抗、高抗和抗病品种。     

Sources of resistance to diseases of sugar beet in related Beta germplasm: II. Soil-borne diseases

Between 580 and 700 accessions of related cultivated and wild species of the genus Beta were assessed for resistance to four soil-borne diseases of sugar beet: two seedling damping-off diseases caused by the fungi Aphanomyces cochlioides and Pythium ultimum and two diseases of more mature plants, Rhizoctonia root and crown rot, caused by the fungus R. solani, and Rhizomania, caused by Beet necrotic yellow vein virus (BNYVV), a furovirus transmitted by the plasmodiophorid Polymyxa betae. Analysis of resistance data (assessed on an international standardised 1–9 scale of Resistance Scores) indicated that the highest levels of resistance ({RS} 2) to A. cochlioides and P. ultimum were to be found amongst accessions of the more distantly related sections Corollinae (93% of accessions tested) and Procumbentes (10%), respectively; although useful levels could also be found in the more closely related, and sexually compatible, section Beta (1–6%). Resistance to Rhizoctonia was also found in section Beta (5–7%), depending on whether field or glasshouse tests were used, but there was little evidence of generally high levels of resistance to Rhizomania among accessions of this section. None of the accessions of sections Corollinae and Procumbentes exhibited any notable resistance to Rhizoctonia. However, all sections Procumbentes and some sections Corollinae (4%) accessions were highly resistant to Rhizomania. Individuals with high levels of resistance to Rhizomania were identified from within some section Beta and Corollinae accessions, in which there was evidence of segregation.     

Sugar beets homozygous for resistance to beet cyst nematode (Heterodera schachtii Schm.), developed from monosomic additions of Beta procumbens to B. vulgaris

Summary Monosomic additions of Beta vulgaris x B. procumbens with resistance to beet cyst nematode (Heterodera schachtii Schm.) were used for the production of resistant diploids through incorporation of the B. procumbens chromosome fragment bearing the resistance gene(s) into one of the sugar beet chromosomes. The heterozygotes obtained accordingly were selfed for producing homozygotes. These homozygotes differed morphologically from commercial sugar beet varieties, but produced reasonable amounts of pollen. Female transmission of resistance was 100%, whereas male transmission, apart from some exceptions, was more than 90%. The number of hypersensitivity reactions to penetrated larvae was related to the degree of susceptibility. Larval development was severely retarded in the resistant plants, preventing most of them to produce cysts. If cysts were formed, their content was considerably less as compared to those in the susceptible plants.     

Identification of resistant sources and DNA markers linked to genomic region conferring dry root rot resistance in chickpea (Cicer arietinum L.)

A set of 520 chickpea germplasm lines was screened under laboratory conditions using blotter paper technique for reaction to dry root rot caused by Rhizoctonia bataticola (Taub.) Butler. The lines PG06102, BG2094 and IC552137 were identified as resistant for dry root rot. Phenotyping the mapping population consisting of 129 F_2:3 progeny derived from the cross L550 × PG06102 during 2013 winter indicated monogenic inheritance of dry root rot resistance. Fifty‐two of 381 simple sequence repeat (SSR) primers polymorphic between the two parents were used to genotype F₂ resistant and susceptible bulks prepared on the basis of reaction of F_2:3 progeny. Four markers differentiated the resistant and susceptible bulks. All the four polymorphic markers were then assayed on the entire F₂ population. Linkage analysis using 129 F₂ plants revealed that two markers ICCM0299 and ICCM0120b were co‐segregating with resistance to dry root rot. These two markers appeared to have additive effects on resistance and could be potentially utilized in dry root resistance breeding programme.     

中国甜菜主要种质资源抗病性鉴定与评价

旨在鉴定与评价不同生态区甜菜主要种质资源的抗病性,为种质资源在抗病育种上科学有效利用提供依据。以甜菜3个生态区的313份主要种质资源为试材,在自然发病条件下,按照甜菜的规范标准,调查种质的发病级数,计算病情指数,划分抗性级别。鉴定出高抗甜菜褐斑病种质资源85份、抗病89份,抗甜菜根腐病种质资源11份,抗甜菜丛根病种质资源3份,抗甜菜白粉病种质资源28份。同时鉴定出兼抗2种病害的种质资源,高抗褐斑病兼抗根腐病种质资源5份、抗褐斑病兼抗根腐病种质资源5份、抗褐斑病兼抗丛根病种质资源2份,高抗褐斑病兼抗白粉病种质资源20份。结果表明,中国甜菜种质资源比较匮乏,除抗褐斑病种质资源外,抗根腐病、丛根病和白粉病的资源不但数量少,而且抗性级别低。     

甜菜抗病品种产生抗性的土壤微生物机理

本研究以甜菜和根腐病为研究对象,分析了不同品种以及不同发病程度甜菜中土壤微生物群落组成、结构以及功能的变化,以期获得抗性品种、发病程度和根际微生物相互间的关系,揭示抗病品种的土壤微生物机制。利用抗病型和感病型两种甜菜品种作为研究对象,并从2种品种中分别选取了发病轻和发病重的甜菜根际土壤样品,通过Illumina MiSeq高通量测序技术对根际土壤细菌和真菌群落进行测定。结果发现：发病较轻的甜菜根际土壤微生物（细菌和真菌）和抗病型甜菜根际土壤真菌的多样性相对较高。NMDS分析表明甜菜品种能显著影响土壤真菌的群落结构,而发病程度则能同时明显改变细菌与真菌的群落结构。在根际微生物富集方面,抗病型甜菜根际富集了假单胞菌(Pseudomonas)、节杆菌(Arthrobacter)、芽孢杆菌(Bacillus)等有益细菌,而感病型甜菜中则富集了较多的病原微生物-尖孢镰刀菌(Fusarium oxysporum)。另外,发病较轻样品中富集了较多的未分类的酸杆菌纲属、芽孢杆菌(Bacillus)、红色杆菌属(Rubrobacter)、未分类的放线菌纲属、链霉菌属(Streptomyces)、类诺卡氏属(Nocardioides)等有益细菌。除此以外,FUNGuild功能预测表明,感病型甜菜和发病较重样品中检测到更多的植物病原菌。抗病品种和健康植株根际微生物虽然在种类上不同,但是其根际均聚集了大量的有益微生物,而感病型品种和发病严重植株则更容易在根围定殖病原微生物。抗病品种的抗病机理之一是植物生长过程中招募了更多的有益细菌作为抵御病原菌的侵染第一道防线。本研究从土壤微生物组的角度对抗病品种抗性机制进行了阐明,充实了抗病品种抗性产生的理论机制。     

Partial resistance of sugarbeet to beet cyst eelworm (Heterodera schachtii Schm.)

Summary Within Beta vulgaris and B. maritima origins some partial resistance to beet cyst eelworm was found which could be raised to a very limited extent by selection. However after the second backcross to commercial sugar beet varieties and successive selection of the inbreds this resistance was lost. It was demonstrated that in the rootsystem of resistant plants as much nematodes penetrate and develop as in susceptible ones but the ratio between males and females is different. It was therefore quite probable that this resistance is polyfactorial and merely recessive.     

基于灰色关联度分析和主成分分析法评估糖用甜菜品种的适应性

为了筛选出最适宜黑龙江哈尔滨地区种植的产质量高并抗根腐病的糖用甜菜品种。2020年在黑龙江省哈尔滨市黑龙江大学呼兰校区试验基地,以21个引种的KWS系列及1个BTS2730糖用甜菜品种（KWS1197为对照）为试材,采用主成分分析和灰色关联度分析法对根产量、含糖率、产糖量和根腐病4个指标进行综合评价。两种方法得出的甜菜品种的排序大致一致;第一主成分根产量的贡献率为69.704%,第二主成分含糖率的贡献率为26.283%,累计贡献率为95.987%,因此能够全面地反映甜菜的产质量性状;最适合本地种植的综合评价值高于对照的6个品种为：KWS0023(0.8231)>KWS0015(0.7685)> KWS6661(0.7511) >KWS9921(0.7103)>KWS0860(0.7097)>BTS2730(0.7065)>CK(0.6823);其他品种的综合评价值低于对照。主成分分析法和灰色关联度分析能够较为全面得分析甜菜品种,得出的结果具有可靠性。     

优异甜菜种质资源筛选评价

旨在筛选评价优异的甜菜种质资源,为在育种上科学利用提供可靠信息支撑。本研究对277份甜菜种质资源的块根产量、含糖率、有害性非糖分以及主要病害抗性等8个主要性状的试验数据,按规范标准进行分组划类,分别对各类型种质资源进行筛选与评价。筛选出块根产量52 t/hm²以上的高产型种质资源37份,含糖率17%以上高糖型88份。其中,高产高糖型4份、高产中糖型的21份、中产高糖型的72份。种质资源中有害性非糖分：钾含量低于3.79 mmol/100 g的25份、钠含量低于2.00 mmol/100 g的96份,α-N低于4.76 mmol/100 g的49份,其中,低钾、低钠含量的4份,低钾、低α-N含量的4份,低钠、低α-N含量的20份,低钾、低钠、低α-N含量的3份。参试种质资源中具有高抗褐斑病兼中抗立枯病的种质资源10份,高抗褐斑病兼抗根腐病的5份,抗褐斑病兼中抗立枯病的14份,抗褐斑病兼抗根腐病的5份、抗褐斑病兼中抗根腐病的3份。结果表明,现有甜菜种质资源中高产型较少、高糖型的比较丰富;抗褐斑病种质资源较多,抗立枯病和抗根腐病的数量少并且抗性级别低。     

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.     

Screening techniques and sources of resistance to root rots and wilts in cool season food legumes

Soilborne, fungal pathogens of cool season food legumes, including seed and seedling blights, rot rots, and wilts are described. Seed and seedling diseases are caused primarily by Pythium and Rhizoctonia spp. The most important fungi causing root rots include Aphanomyces euteiches, Fusarium solani, Pythium spp., Sclerotium rolfsii, and Macrophomina phaseolina. Wilt is caused primarily by various host-specific forms of Fusarium oxysporum. This paper discusses these diseases and screening procedures that emphasize standardization of inoculum levels, maintenance of virulent pathogen cultures, inoculum growth media, environmental conditions, and host plant age. Sources of resistance to these diseases are discussed.     

Molecular mapping of a gene conferring resistance to Aphanomyces root rot (black root) in sugar beet (<Emphasis Type="Italic">Beta vulgaris</Emphasis> L.)

Caused by Aphanomyces cochlioides Drechsler, Aphanomyces root rot is a serious disease of sugar beet (Beta vulgaris L.), for which sources of resistance are scarce. To identify the segregation pattern of the rare resistance trait found in Japanese sugar beet line ‘NK-310mm-O’, F₁ and BC₁F₂ seedings, drawn from a cross between ‘NK-310mm-O’ and susceptible line ‘NK-184mm-O’, were inoculated with zoospores and their survival evaluated in the greenhouse. Resistance segregation followed was that of a single dominant gene, which was designated Acr1 (Aphanomyces cochlioides resistance 1). Molecular markers tightly linked to Acr1 were identified by bulked segregant analysis of two BC₁F₂ populations. Fourteen AFLP markers linked to Acr1 were identified, the closest located within ±3.3 cM. Three F₅ lines and two BC₂F₁ lines, selected on the basis of their Acr1-AFLP markers, were tested for their resistance to Aphanomyces root rot in a highly infested field. Results indicated that Acr1 conferred significant resistance to Aphanomyces root rot at the field level. Based on its linkage with CAPS marker tk, a representative marker for chromosome III, Acr1 was located on this chromosome. The clear linkage between tk and Rhizomania resistance trait Rz1, suggests the clustering of major disease resistance genes on chromosome III.     

草地早熟禾对立枯丝核菌侵染的生理响应

为探索立枯丝核菌(Rhizoctonia solani)侵染后草地早熟禾(Poa pratensis L.)的生理响应,以2个对褐斑病抗性不同的草地早熟禾品种‘午夜’和‘蓝孔雀’为材料,在接种立枯丝核菌后,统计了草坪草的发病率和病情指数,并测定分析不同时间点草坪草叶片内的相对电导率值、丙二醛含量、脯氨酸含量、叶绿素含量等生理指标的变化规律。结果表明：与接种前相比,接种后的草叶随着时间的推移,其病情指数、相对电导率、丙二醛和脯氨酸含量都显著增大,而叶绿素含量显著降低。其中,易感病品种‘蓝孔雀’的病情指数、相对电导率、丙二醛和脯氨酸含量的增幅均高于抗病品种‘午夜’,而且前者叶绿素含量的降幅也大于后者。相关分析表明,丙二醛含量、脯氨酸含量和相对电导率与病情指数的变化呈显著正相关,叶绿素含量与病情指数呈显著负相关。     

甜菜抗丛根病育种研究进展

为了研究甜菜抗丛根病育种的抗性基因来源以及作用特点,笔者归纳了甜菜丛根病抗性的来源,包括Alba抗性源、Rizor和Holly抗性源、WB42抗性源、WB41和WB258抗性源以及R36抗性源,并介绍了5种甜菜丛根病的抗性基因特点。另外,对甜菜抗丛根病资源的育成方法进行了总结,包括回交、杂交和轮回选择。通过这些方法将携带抗丛根病资源的甜菜整合至适应性高的甜菜种质资源上,从而获得理想的抗性资源。针对甜菜抗丛根病育种过程中培育和选择甜菜丛根病抗性基因型的过程缓慢、费力且昂贵,以及具有抗性基因品种的抗性易被破坏的情况,现提出如下解决策略：（1）通过分子标记来选择理想的甜菜新品种;（2）不断鉴定新的抗性基因,并将其整合到甜菜的基因库中;（3）开发新抗源,通过鉴定其他基因座上与先前发现的抗性基因的等位基因,为释放具有足够抗性的品种提供机会。抗甜菜丛根病育种的下一个挑战将会是将新的抗性基因导入育种材料,并通过将不同基因座上的多个抗性基因聚合到育种系中进行组合,以最大限度地提高甜菜对丛根病的抗性水平。     

A simple method based on laboratory inoculum and field inoculum for evaluating potato resistance to black scurf caused by Rhizoctonia solani

A two-step method was developed to evaluate potato resistance to black scurf caused by Rhizoctonia solani. Tuber piece inoculum was first conducted in the laboratory, which was also first reported in this study. After inoculation with pathogen discs and culture for 48 h, the necrotic spots on the inoculated potato pieces were generated and measured by the crossing method. Further evaluation was conducted through field experiments using a wheat bran inoculum method. The wheat bran inoculum was placed into the pit dispersedly and surrounded seed tubers. Each cultivar or line was subjected to five treatments of 0-, 2-, 3-, 4-, and 5-g soil inoculum. The results showed that 2–4 g of wheat bran inoculum was the optimum for identifying tuber black scurf resistance. The laboratory scores positively correlated with the incidence and severity of black scurf in the field. According to the results in the laboratory, relatively resistant cultivars could be selected for further estimation of tuber black scurf resistance in field experiments. It is a practical and effective screening method for rapid identification of resistant potato germplasm, which can reduce workload in the field, shorten time required for identification.     

高抗褐斑病性和含糖率甜菜品种田间鉴定

为选择抗褐斑病性高、含糖率高的优良甜菜品种用于生产,提高经济效益。2015—2016 年,采用田间试验自然发病鉴定方法,15 个品种为处理,随机区组排列,4 次重复,对商用甜菜品种的褐斑病病情指数、含糖率进行调查测定。统计分析结果表明：不同甜菜品种间抗褐斑病性差异极显著,甜菜品种的抗褐斑病性不稳定,不同年份间差异较大。品种间含糖率差异极显著。同一品种不同年份含糖率存在差异,抗褐斑病性高并相对稳定、含糖率高的甜菜品种为‘KWS1197’,2 年的平均病情指数为18.34,平均含糖率为14.01%。