Genetic Relationships Among Four Pepper Genotypes Resistant to Phytophthora capsici

According to our previous investigations, resistance to Phytophthora capsid in Capsicum annuum genotypes, ‘Line 29’, ‘PI201232’, ‘PI201234’ and Serrano Criollo de Morelos 334 (‘SCM334’), seems to be controlled by three genes. In order to determine the genie relationships between these four sources of resistance, three experiments were conducted which included the four genotypes, their F₁s, F₂s, F3s and BC₁ generations together with the susceptible pepper genotype ‘Morron INI A 224’. Inoculations were made, when plants had 4—6 leaves, by irrigating the culture substrate with a zoospore suspension of P. capsici isolate ‘Bl’. Though the four genotypes showed percentages of resistance close to a 100%, none of them actually reached this level in the three experiments. ‘SCM334’ was the most resistant genotype, transmitting a high level of resistance to its F₁, F₂ and BQ generations. ‘Line 29’ was more resistant than ‘PI201232’ and ‘PI201234’. However, the F₁ F₂ and BQ generations of these three lines showed similar degrees of resistance. The four genotypes seem to have one of the three genes postulated for their resistance in common. All genes displayed a similar level of resistance, except the specific genes of ‘SCM334’, the effect of which was slightly higher. Several working procedures are suggested for breeding programmes.     

不同辣椒材料对疫病的抗性研究

为筛选抗性种质资源,进行抗性品种选育和推广种植,利用苗期游动孢子灌根法对206份江西省地方辣椒材料进行抗病性鉴定,并初步分析不同时期材料的发病情况。研究结果表明,不同材料间的抗病性存在较为明显的差异,无论是病株率还是病情指数均有显著差异性,病株率在0～30.0%、30.1%～70.0%、70.1%～100%之间的材料分别为21、32、153份,占供试材料的10.19%、15.53%、74.27%;表现为高抗、抗病、中抗和感病的材料分别有15、17、23、151份,占供试材料的7.28%、8.25%、11.17%和73.30%;鉴定出的‘赣椒18号’‘、赣椒35号’、C047、WY38、CM334和A204等15份高抗材料是很好的抗疫病资源。分析供试材料在2次调查结果中的发病情况可知,材料间的发病速度(侵入和扩展)存在显著差异性。     

Interactions in the pepper-Phytophthora capsici system

To analyse interactions in the Phytophthora capsici-pepper (Capsicum annuum L.) system, three pepper lines (‘Morrón IN1A 224’, ‘PI201232’ and ‘SCM 334’) with various degrees of resistance to P. capsici and three P. capsici isolates with increasing aggressiveness were used. Inoculations were made when plants had 4–6 leaves, either by irrigating the culture substrate with P. capsici zoospore suspensions at increasing concentrations of up to 3 × 10⁵/ml, or by adding mycelial suspension. P. capsici isolate ‘Ca’ was the least aggressive, followd by ‘8303’ and ‘Bl’. The logistic function was appropriate to describing cumulative mortalities of pepper seedlings inoculated with different P. capsici zoo-spore concentrations. The inoculum concentration required for 50% mortality classified pepper lines as susceptible (‘Morrón INIA224’) and resistant (‘PI201232’ and ‘SCM334’), the former being less resistant than the latter. Isolates ‘Ca’ and ‘8303’ caused very different mortalities both with zoospore and mycelial inoculations, which allowed them to be classified as belonging to two different vertical pathotypes (PO and PI). Isolate ‘Bl’ acted similarly to isolate ‘8303’ when inoculated on ‘Morrón’ and ‘SCM334’, but its behaviour was dependent on zoospore concentration when inoculated on ‘PI201232’. It also proved to be similar to ‘Ca’ using mycelial inoculation. As the case of ‘Bl’ would not be an exception among P. capsici isolates, the two vertical pathotypes PI and PO would be the higher (isolate ‘8303’) and lower (isolate ‘Ca’) limits of a range of aggressiveness. Although there are cases of host-parasite interaction, absence of interaction seems to be the rule.     

水杨酸诱导辣椒抗疫病生化机制的研究

对水杨酸（SA）处理及其挑战接种辣椒疫霉菌的4个抗性不同的辣椒品系体内木质素含量和多酚氧化酶（PPO）、过氧化物酶（POD）、苯丙氨酸解氨酶（PAL）活性的测定结果表明：SA诱导的供试品系与未经诱导的相同品系相比,木质素含量增高,PPO、POD、PAL活性增强,其中木质素的含量与抗病性呈显著正相关;挑战接种后两者相比,SA处理的供试材料的木质素含量和PPO、POD、PAL活性的增强幅度更大,其中抗、耐病品系的增加早且增加幅度高于感病、高感品系。值得注意的是,经SA诱导的各供试品系接种后木质素含量与抗病性也呈显著相关。木质素含量与相关酶活性的上述变化趋势与SA对各品系的诱抗效果完全吻合。     

氨基寡糖素田间防治辣椒疫病及体外抑菌试验

研究了氨基寡糖素田间防治辣椒疫病和体外对辣椒疫霉(Phytophthoracapsici)的抑菌作用,为其在生产上的应用提供依据。田间喷雾使用氨基寡糖素对辣椒疫病有一定的防效,40mg/L氨基寡糖素对辣椒疫病的防效高达73.2%。采用生长速率法在马铃薯葡萄糖琼脂(PDA)上研究了氨基寡糖素体外对辣椒疫霉菌丝生长的影响,结果表明氨基寡糖素可以抑制辣椒疫霉的菌丝生长,有效中浓度EC50为100mg/L。在无菌水中氨基寡糖素可以抑制辣椒疫霉新生菌丝上孢子囊的形成以及静止孢的萌发,有效中浓度EC50分别为0.64mg/L和41.84mg/L。     

Inheritance of resistance to anthracnose (Colletotrichum capsici) at seedling and fruiting stages in chili pepper (Capsicum spp.)

Inheritance of resistance to anthracnose at fruiting and seedling stages was studied in two F₂ populations from a cross between Capsicum annuum cv. ‘Bangchang’ and Capsicum chinense‘PBC932’. The first F₂ was used to study anthracnose resistance at fruiting stage on mature green and ripe red fruit, and the second F₂ was used to study the resistance at both seedling and fruiting stages. Fruit inoculation was performed on detached fruit using a microinjector. Disease severity was assessed on a 0–9 scale at 7 days after inoculation. Seedling inoculation was performed using a drop method on detached leaves of 4‐week‐old seedlings. Disease severity was assessed on a 1–9 scale at 3 days after inoculation. The distribution of the disease scores of green and red fruit, and seedlings in both F₂ populations suggested a single gene model for each trait. Three different recessive genes were responsible for the three resistances from this cross. Linkage analysis suggested that the resistances at green and red fruit were linked (recombination frequency 0.25), and that the seedling resistance was not linked to the fruit resistances.     

Inheritance of resistance to anthracnose caused by Colletotrichum capsici in Capsicum

Inheritance of resistance to anthracnose caused by Colletotrichum capsici (Syd.) Butler & Bisby was studied in interspecific Capsicum populations derived from a cross between a Thai elite cultivar Capsicum annuum L.‘Bangchang’ and a resistant line C. chinense Jacq.‘PBC932′. The resistance was assessed by measuring lesion area per fruit area (LFA) on detached chili fruits, using a laboratory‐based injection inoculation. Nil symptoms resembling the resistant parent ‘PBC932’ were also identified in the progeny F₂ and BC1 populations. Segregation of resistance (nil LFA) and susceptibility in the F₂ fitted a 1: 3 Mendelian ratio, indicating that resistance was responsible by a single recessive gene. The segregation of the trait in the testcrosses in both BC1s also confirmed the 1: 3 gene segregating model as found in the F₂.     

辣椒疫霉菌粗毒素对辣椒种子发芽及幼苗生长特性的影响

辣椒疫霉菌（Phytophthora capsici）可对寄主植物产生致病性的毒素,使辣椒产生类似病原菌侵染引起的症状。将辣椒疫病病原菌粗毒素按10％,20％,30％,40％,50％,60％,70％,80％,90％,100％的体积比浓度,分别作用于未萌发和已萌发的辣椒种子,结果表明,辣椒疫霉菌粗毒素对辣椒种子发芽指数、辣椒幼苗株高、下胚轴长以及辣椒胚根的主根长、侧根长、侧根。数具有明显的抑制作用。通过主、侧根抑制率的比较,在同一浓度下,粗毒素对侧根的抑制作用大于主根。     

疫病病菌侵染后辣椒幼苗体内保护酶活性的变化

痰霉菌侵染后辣椒幼苗叶片和根茎组织中PPO、POD和PAL活性发生变化。试验表明：除感病品种根茎部固有的POD活性较高以外，抗(耐)病辣椒品种幼苗叶片的PPO、POD和PAL及根茎部PPO和PAL活性高于感病品种。痰霉菌侵染后，仅根茎部PPO活性略有下降，各辣椒品种幼苗叶片和根茎组织PPO、POD和PAL均在接种后一度显著高于对照。抗(耐)病辣椒品种幼苗根茎部PAL活性接种4d升幅大且早，抗(耐)病品种体内固有的PP0、PCD和PAL活性高，在辣椒抗疫霉菌反应中起了重要作用。     

Inheritance of adult-plant resistance to Phytophthora capsici in pepper

Summary Inheritance studies were conducted to determine the genetic basis of adult-plant resistance in pepper (Capsicum annuum L.) to Phytophthora capsici. F₁, backcrosses and F₂ populations were developed using the resistant parent Criollo de Morellos 334 and susceptible parents Agronômico 10-G and Yolo Wonder. Pepper plants, at 36 days post-emergence, were inoculated near the base of the stem with an inoculum suspension of 5×10⁴ zoospores/ml. Segregation ratios in the F₂ generation of 13 resistant to 3 susceptible plants fit a 2-gene model for resistance with dominant and recessive epistasis.     

辣椒疫霉菌菌种保存及致病性研究

在辣椒疫病抗性材料鉴定和疫霉菌生理小种测定等工作中, 常因菌种在室内连续多代转接培养导致菌株的致病力下降, 影响测定结果的准确性和一致性。针对辣椒疫霉菌的长时间保存这一问题, 通过对 4个辣椒疫霉菌株的长时间无菌水保存和常规固体培养基斜面连续转接保存方法进行试验对比,定期对菌株培养性状和致病力进行观察和测定。试验结果表明： 在 16℃条件下, 无菌水中保存的辣椒疫霉菌株在 24个月后仍有活力, 菌株培养性状没有发生变化, 致病性也没有明显衰退。该菌种保存方法制作简单、 经济、 可靠, 适用于辣椒疫霉菌种的长期保存。     

甜瓜种质资源对疫病的抗性鉴定研究

旨在通过苗期人工接种鉴定,筛选高抗疫病的甜瓜材料,为甜瓜抗病育种以及抗疫病遗传学研究奠定基础。本研究采用灌根接种法,向植株根茎部土壤注入浓度为1×106个/mL的游动孢子悬浮液,对收集到的166份甜瓜材料进行了抗性鉴定,这些材料中有薄皮类型甜瓜41份,厚皮类型甜瓜104份,野生材料15份,近缘种6份。最终筛选到4份免疫材料,25份高抗材料。在供试的厚皮类型甜瓜中有66%的材料抗性水平在中抗及以上,而薄皮类型中有98%的抗性水平是感病或高感。通过相关性分析,发现厚皮类型甜瓜对疫病的抗性要显著高于薄皮类型甜瓜。通过对不同材料的果实性状进行调查,发现在免疫和高抗材料中有14份单瓜重达到1 kg以上,有8份果实可溶性固形物含量在10%以上。这些兼具高抗、高产和高可溶性固形物的材料,可用于抗病基因的定位、克隆等遗传学研究,也可作为育种亲本将抗病基因转移到农艺性状优良但抗性差的甜瓜品种中。     

辣椒疫霉菌ITS及β-tubulin的扩增和序列分析

为了探索湖南省不同地区辣椒疫霉的系统发育关系与遗传多样性,从湖南芷江、宁乡等地采集辣椒疫病病株,经过分离和离体叶片接种,得到2株对辣椒叶片有强致病性的辣椒疫霉菌(Phytophthora capsici)菌株。利用PCR技术对所分离的2个菌株进行ITS以及β-tubulin扩增并测序,将测序结果与国内外报道的相关序列构建系统发育树,结果表明,离体叶片接种法能够对辣椒疫霉菌有一个初步的鉴定,PCR扩增的ITS序列长度为770 bp左右,β-tubulin序列长度为780 bp左右,ITS序列以及β-tubulin序列构建的系统发育树表明湖南省内辣椒疫霉的亲缘性比较近,与国外的亲缘性比较远,说明了不同地区疫霉菌株的亲缘性与地理来源有一定的相关性。     

辣椒砧木种质资源对疫病的抗性及遗传初步分析

以从国外引进的辣椒砧木种质资源为研究对象,采用游动孢子灌根法对幼苗进行疫病抗性鉴定;选择不同抗性水平且抗性表型稳定的5份砧木种质进行完全双列杂交,鉴定F1代对疫病的抗性,以病情指数为表型指标分析抗性杂种优势,通过Griffing法分析抗病性的配合力和遗传参数,探讨砧木种质抗病性状的数量遗传特点.结果表明:15份供试辣椒砧木种质中,3份种质对疫病表现中抗,其余种质均表现感病;20个F1杂交组合中,有2个杂交组合对疫病表现中抗,有3个组合表现杂种优势;辣椒砧木种质抗疫病性状符合“加性-显性-上位性”数量遗传模型,且加性效应占主导地位,同时可能受细胞质基因的影响,表现核质互作效应.本研究筛选出3份辣椒砧木种质和2个F1杂交组合对疫病表现中抗,其中种质D15可作为抗疫病育种骨干亲本,研究结果可为合理利用辣椒砧木种质开展抗疫病育种提供参考.     

辣椒疫病生防菌株鉴定及控病机理研究

为筛选出对辣椒疫病菌具有较强抑制活性的生防菌株,达到控制辣椒疫病的目的,从辣椒种植区土壤中分离大量微生物,采用平板对峙法筛选对辣椒疫病菌具有较强抑制作用的生防菌株XD6,通过常规方法和16SrDNA确定其分类地位,并测定相关酶活性变化。试验分离得到的XD6菌株,平板对峙试验防治效果高达80%~100%,初步鉴定为多粘芽孢杆菌(Paenibacillus polymyxa)。酶活测定显示,生防菌株XD6的β-1,3-葡聚糖酶和纤维素酶活性都存在显著变化,且接种XD6菌株的辣椒植株PAL活性、PPO活性、POD活性均明显升高。菌株XD6对辣椒疫病具有较高防效,其生防因子可作用于辣椒疫病菌细胞壁,并诱导辣椒自身防御酶系的增强,具有良好的研究前景。     

Resistance to Phytophthora infestans in Potato Genotypes Originating from Wild Species

The reaction to P. infestans in the foliage and tubers of potato was evaluated in resistant genotypes and in their progeny. The genotypes originated from wild species known to carry genes for specific resistance. Evidence was obtained that three types of resistance to P. infestans were presented in the genotypes: (1) Resistance due to major, dominant genes was predominant. The presence of these genes was expressed in both foliage and tubers and did not influence maturity. Some of the genes were R genes, providing specific resistance, but for others this could not be determined as they were effective against the fungus inoculum with the highest virulence available: 1.2.3.4.5.7.10.11. (2) Resistance was present, associated with late maturity. (3) Tuber resistance was present, expressed after inoculation of whole tubers but not after inoculation of tuber slices. Testing of the progeny was essential for the determination of the type of resistance present in the genotypes evaluated.     

利用花药培养技术创制加工型辣椒抗疫病新种质

为培育抗疫病加工型辣椒品种,本研究利用花药培养与分子标记辅助选择(molecular assisted selection,MAS)相结合的方法将抗疫病材料PPTC54和PI201234的抗性基因导入优良自交系939、812、83-3、968和花培双单倍体(doubled haploid, DH) H12-11,进行抗疫病新种质创制的研究。结果表明,在5个加工型辣椒基因型中以(PPTC54×83-3) F1诱导出的胚状体成苗率最高,可达87.1%;流式细胞仪测定36个花培再生株的DNA含量分布,其中单倍体株数占80.6%;经PCR检测,筛选出14个抗疫病DH系,且聚合抗疫病基因的各株系间农艺性状存在差异。本研究说明结合花药培养与MAS可快速创制特异育种新种质,加速育种进程。     

Gene action in the resistance of peppers (Capsicum annuum) to Phytophthora stem blight (Phytophthora capsici L.)

Summary The variation among single and three-way crosses derived from seven lines of pepper (Capsicum annuum) selected for resistance to Phytophthora capsici was analyzed into statistical-genetic components due to general and specific combining abilities and additive-by-additive epistasis. Indirect evidence of higher order epistasis was also estimated. Epistasis was a principal source of variation. It was correlated with the level of aggressiveness of the pathogen. A breeding method such as recurrent selection seems to be the most appropriate to improve resistance to the fungus.     

不同地区辣椒疫霉菌生理小种的鉴定和生物学特性的研究

为了确定不同地区采集的辣椒疫病病原菌的菌种类型及其生理小种类型,首先利用形态学和分子生物学的手段对广东、山西和内蒙古等地的辣椒疫病的病原菌进行了鉴定,结果表明,所分离到的7株病原菌均为辣椒疫霉菌。通过比较不同地区分离到的疫霉菌生物学特性,发现不同地区辣椒疫霉菌在菌落形态、孢子囊的数量、生长速度等方面存在显著的差异。利用国际通用的鉴别寄主,对分离到的7个辣椒疫霉菌进行生理小种的鉴定,结果表明,来源于广东的辣椒疫霉菌P1是2号生理小种,而来源于山西和内蒙古的6株辣椒疫霉菌P2~P7都是3号生理小种。目前,在山西和内蒙古地区并没有关于辣椒疫霉菌生理小种鉴定的任何报道,确定了这2个地区的辣椒疫霉菌的优势生理小种为3号生理小种。     

Breeding transgressive lines of pepper for resistance to Phytophthora capsici in a recurrent selection system

Summary In order to increase the resistance level of pepper (Capsicum annuum) to Phytophthora capsici the main sources of genetic resistance were intercrossed. The parents included 7 varieties from different origins of partial resistance to P. capsici and to other soilborne pathogens. Two cycles of recurrent selection were performed by 7 plant breeders who screened the plants at two developmental stages with various isolates and at two temperatures. Analysis of data revealed a strong influence of the testing conditions on the evaluation of the plants. The level of resistance of the progenies further depended on the pathogen isolate used to screen the plants. Improvement of the whole population was more significant during the first cycle and lines with enhanced level of resistance as compared to the parents i.e. transgressive lines were fixed from the two cycles. These lines were resistant to much higher inoculum concentrations than the original parents and the expression of resistance was stable at high temperature (32°C). These new sources of polygenic resistance were included into a new breeding program to improve the agronomic characters.