Chromosome localisation of genes for resistance to Heterodera schachtii, Cercospora beticola and Polymyxa betae using sets of Beta procumbens and B. patellaris derived monosomic additions in B. vulgaris

Beet cyst nematodes (BCN, Heterodera schachtii), Cercospora beticola, and rhizomania, caused by the beet necrotic yellow vein virus (BNYVV) and vectored by the soil-borne fungus Polymyxa betae, are the most serious diseases of sugar beet ( Beta vulgaris subsp. vulgaris). The wild Beta species of section Procumbentes are known to be completely resistant to H. schachtii, C. beticola and P. betae. Alien monosomic additions (2n=19), plants of cultivated beet (2n=18) carrying different individual chromosomes of B. procumbens (2n=18) or B. patellaris (2n=36), were tested in greenhouse experiments for resistance to these pathogens. Gene(s) conferring full resistance to the beet cyst nematode in B. patellaris are located on chromosome 1.1, and the other tested chromosomes of B. patellaris are not involved in the expression of resistance. Artificial inoculation under greenhouse conditions, with in vitro produced inoculum of C. beticola and spot-percentage rating of the disease intensity, showed that the high level of resistance that was observed in the wild species B. procumbens and B. patellaris was not found in any of the monosomic additions tested. It was suggested that genes on various chromosomes of the wild species are needed to express full resistance, and that the chromosomes of group 7 of B. patellaris and chromosome 7 of B. procumbens have the largest effect. The greenhouse tests for resistance to P. betae in B. patellaris derived monosomic additions showed that the addition families of group 4.1 have a strong partial resistance, while the addition families of group 8.1 appeared to be completely resistant to the pathogen. Resistance to P. betae in the two wild species as well as in the two resistant addition types did not exclude infection with BNYVV, but resulted in a considerable reduction of the virus concentration. It was concluded that resistance to the vector would complement virus resistance, and may provide a more effective and durable control of rhizomania. This revised version was published online in July 2006 with corrections to the Cover Date.     

郑196耐大豆胞囊线虫病研究及耐病机理初探

郑196是从黄淮海大豆产区选育的优良大豆品种。该品种在SCN2病圃中鼓粒情况良好;2017年,郑196在黄淮海9个试点的产量与在SCN2病圃中产量相比较,差异不显著,说明该品种具有耐SCN病的特性;利用荧光定量PCR,进一步分析来自Rhg1/Rhg4位点的4个SCN-抗性基因(Glyma18g02580, Glyma18g02590, Glyma18g02610, SHMT)在郑196及其他不同抗性水平大豆中的表达,结果表明:在受到SCN2侵染的0～25 d,4个SCN-抗性基因在抗病材料中的表达量均持续提升;在受到SCN2侵染的10 d/15 d,这4个SCN-抗性基因在郑196及感病材料中表达量达到最高点,之后表达量下降,该结果表明,郑196的耐SCN病机理不同于SCN抗性基因在抗病材料中的抗病机理,其耐病性不是由SCN抗性基因单独调控的,有可能存在其特有的耐病通路或是由抗性基因与耐病基因共同调控其耐病机制。本研究可对抗SCN种质资源创新和抵御SCN策略提供参考依据。     

Identification and resistant characterization of legume sources against Meloidogyne incognita

Root-knot nematodes(RKNs; Meloidogyne spp.) are becoming a serious problem in legume production. This study identified Vigna genotypes exhibiting resistance to M. incognita(RKN) and characterized the modes of the resistance to M. incognita. In total, 279 accessions from 21 Vigna species were screened for resistance based on a galling index(GI) and an egg mass index(EI). Seven accessions were highly resistant to RKN with GI≤25, namely JP74716(V. mungo var. mungo; cultivated black gram), JP107881(V. nepalensis), JP229392(V. radiata var. sublobata; wild mungbean), Aus TRCF118141(V. unguiculata subsp. unguiculata; cultivated cowpea), Aus TRCF306385(V. unguiculata subsp. unguiculata), Aus TRCF322090(V. vexillata var. vexillata; wild zombi pea) and JP235929(V. vexillata var. vexillata). JP229392 and Aus TRCF322090 were the most resistant accessions having EI values of 18.74 and 1.88, respectively. Continuous culture of M. incognita on both JP229392 and Aus TRCF322090 resulted in a weakness in pathogenic ability for this RKN. The resistance in JP229392 and Aus TRCF322090 to RKN appeared to be antibiosis that was associated with reduced nematode penetration, retardation of nematode development and impeding giant cell formation. The Vigna germplasm resistance to RKN identified in this study could be utilized as gene sources for the development of RKN-resistant Vigna cultivars.     

莱州湾海域小刀蛏(Cultellus attenuatus)繁殖生物学研究

2013年4月–2014年3月,采用组织学和实验生态学方法研究了山东莱州湾海域小刀蛏(Cultellus attenuatus)的性腺发育、生殖周期、胚胎及幼虫发育。结果显示,在繁殖季节,小刀蛏性别可通过性腺颜色区分,雌性为白色,雄性为黄色;1个生殖周期为1年,性腺发育经历增殖期、生长期、成熟期、排放期和休止期5个阶段;莱州湾繁殖期为6月中旬–7月上旬。小刀蛏受精卵卵径为50–55 μm,在水温26℃、盐度28的条件下,经24 h发育至D形幼虫,10 d后幼虫发育变态为稚贝。对莱州湾小刀蛏繁殖生物学的研究,可为该海域小刀蛏的人工育苗和增养殖提供科学依据。     

Development,Survival and Availability of Gastrointestinal Nematodes of Sheep and Pastures in a Semi-arid Area of Kajiado District of Kenya

A study was carried out on a ranch in the semi-arid area of Kajiado District in Kenya during the period July 2000 to June 2001 to determine the seasonal patterns of development and survival of gastrointestinal nematodes of sheep on pastures. A series of plots were contaminated with sheep faeces every month and pasture samples were collected weekly for the recovery and identification of larvae. The availability of infective larvae on naturally contaminated pastures was also monitored on the paddocks grazed by sheep and around the night pen and the watering point every month from July 2000 to June 2001. The results from the examination of the pasture samples indicated that rainfall distribution was the major factor governing the development and survival of the pre-parasitic stages. No parasitic larvae were detected from the plots contaminated during the dry months from July to October 2000, but development and translocation of infective larvae on pastures occurred on plots contaminated during the rainy seasons and soon after when relatively high moisture was present in the herbage (November 2000 to June 2001). During this period, peak larval counts occurred between the first and the second week post contamination, then declined to undetectable levels between week 4 and 16 post contamination. The lack of development of infective larvae during the dry season and the relatively rapid decline of their population during the wet season presents an opportunity for the use of pasture spelling as a means of helminth control in the study area. The availability of infective larvae on naturally contaminated pastures, around the night pen and around the watering point also followed the rainfall distribution pattern. Infective larvae were consistently recovered around the watering point throughout the study period. This indicated that the point is an important source of infection for sheep, especially during the dry season when other pastures are non-infective.     

Managing anthelmintic resistance: Modelling strategic use of a new anthelmintic class to slow the development of resistance to existing classes

AIM: To test the hypothesis that a single strategic treatment with a new class of anthelmintic could slow the development of resistance to existing classes of anthelmintic.

METHODS: An existing model was used to simulate nematode parasite dynamics and the development of anthelmintic resistance. Variations on a five-drench preventive programme of treatments for lambs, in which either zero, the first, third or fifth treatment was substituted with a different class of drug, were compared for the time to reach treatment failure (defined as efficacy <95%). The sensitivity to variations in the death rate of adult worms, that varied from 1 to 5%, and the dominance of resistance genes were also assessed.

RESULTS: Replacing one of the five treatments with a different class of anthelmintic almost always slowed the development of resistance, and was never worse than using the same drug for all treatments. Further, there were large differences in the relative time to treatment failure depending on which treatment was substituted. Changing the first treatment always had the least benefit, whereas changing the fifth treatment always had the greatest. This pattern was independent of the daily death rate of adult worms, and was not influenced by the dominance of resistance under treatment.

CONCLUSIONS: The results indicated that strategic substitution of a single treatment with a new class of anthelmintic, at the end of a series of preventive treatments to lambs using an existing class, could slow the further development of resistance to the latter. This strategic use of a new anthelmintic class has the potential to greatly extend the life of existing anthelmintics if these are still effective.     

Ontogeny of the stomach in yellow catfish (Pelteobagrus fulvidraco): detection and quantifictation of pepsinogen and H+/K+‐ATPase gene expression

Yellow catfish (Pelteobagrus fulvidraco) is an important commercial species with high aquaculture potential in China. To better understand the process of digestive functioning of gastric gland development during the larval from 1 dph (day post‐hatching) to 30 dph, real‐time PCR was used to detect and quantify the pepsinogen and H⁺/K⁺‐ATPase gene expression in P. fulvidraco. These data were also compared with the adult situation. The results showed that the expression of pepsinogen and H⁺/K⁺‐ATPase genes in P. fulvidraco larvae both started at 1 dph, though the expression level was very low until 3 dph. The quantification of pepsinogen gene expression increased significantly from 4 to 8 dph, increased fluctuantly from 8 to 23 dph and rose sharply from 23 to 30 dph. In comparison with adult fish, there were no significant differences with larvae at 5 and 23 dph. However, data of 10 and 30 dph larvae were obviously higher than those of adult group. H⁺/K⁺‐ATPase gene expression increased linearly from 1 to 30 dph. However, it was significantly lower than that of adult. The results show that P. fulvidraco larvae have an earlier functional stomach, though the function of the stomach is still not perfect. There is a gradual acidification environment within the stomach during the P. fulvidraco larvae development. Based on these results, we suggest that the weaning time for P. fulvidraco larvae would be much better after 23 dph.     

猪蛔虫感染期幼虫抑制消减cDNA文库的构建

为筛选与线虫感染性相关的基因,本研究以猪蛔虫为对象,构建猪蛔虫感染期幼虫差异表达消减cDNA文库,为研究线虫期特异性发育的分子机制奠定基础。分别提取感染期幼虫和其它各期幼虫及成虫的总RNA,纯化mRNA后,采用Clontech公司PCR-selectTM试剂盒进行反转录合成cDNA并进行抑制消减杂交(SSH),构建猪蛔虫感染期幼虫差异表达的消减cDNA文库,并采用Southern斑点杂交进行消减效率的检测。随机从文库中抽取45个克隆进行测序及在线BLAST分析。试验结果表明,感染期幼虫差异表达的消减cDNA文库具有较强的特异性;在得到的41个表达序列标签(ESTs)中,有40个ESTs与已报道的基因有较高的相似性,主要代表猪蛔虫第三期幼虫基因和成虫头部基因,有1个cDNA片段可能代表新基因。猪蛔虫感染期幼虫差异表达消减cDNA文库的成功构建,为进一步研究幼虫发育差异表达基因的功能奠定了基础。     

盐度对真鲷受精卵发育及仔稚鱼生长的影响

在室内水池以溢水法收集真鲷 (Pagrosomusmajor)受精卵 ,并置于不同盐度海水中静水孵化 ,孵出仔鱼以流水培育。设置盐度梯度为 17.0、2 2 .0、2 7.0及自然盐度 3 3 .0 ,水温分别为 19.5～ 2 1.5℃ (孵化 )、2 0 .5～ 2 3 .5℃(培育 )。结果表明∶①真鲷受精卵在自然盐度海水中发育很好 ,孵化率较高 ( 85 .0 %～ 97.0 % ) ,畸形率较低 ( 3 .2 %～ 2 3 .7% ) ;在低盐度条件下静水孵化 ,发育不好 ,孵化率为 4 8.0 %～ 1.0 % ,且盐度越低 ,孵化率也越低。充气孵化时 ,盐度低至 17.0 ,孵化率有时也可达 97.1% ,但畸形率高达 85 .7%～ 10 0 %。②孵化仔鱼中有 2类畸形 ,第 1类畸形表现为身体或尾部弯曲 ,油球位于卵黄囊中部或前部 ,油球数多于 1个 ,盐度越低 ,畸形率越高 ;第 2类畸形表现为仔鱼膜膨大、围心腔扩大 ,这类畸形只出现于低盐度条件下 ,盐度越低 ,畸形率越高 ,且往往与第 1类畸形症状并发。③不同盐度对真鲷仔稚鱼的生长速度没有显著影响 ,各盐度条件仔稚鱼经 3 1d的培养 ,其平均全长为 19.6～ 2 0 .7mm( P >0 .0 5 ) ,没有显著的差别。④海水盐度越低 ,仔稚鱼的存活率越高 ,各组的平均存活率为 ,盐度17.0时 ,3 6.3 % ;盐度 2 2 .0时 ,3 6.2 % ;盐度 2 7.0时 ,2 7.1% ;盐度 3 3 .0时 ,16.3 %。     

基于DNA条形码技术的泰国番石榴中实蝇幼虫分子鉴定研究

实蝇是世界重要的检疫性害虫之一,对果蔬生产及其国际贸易具有很大的影响。而以线粒体DNA细胞色素C氧化酶亚单位I(mtDNACOI)基因的部分序列作为实蝇的DNA条形码能减少对实蝇成虫形态特征的依赖,可检测其任何虫态的样品,有助于实蝇样品的快速鉴定。本研究采用DNA条形码技术,针对采自泰国四色菊市番石榴烂果中的5头实蝇幼虫进行COI扩增测序,与生命条形码数据库(BOLD)中的序列进行比对并利用PAUP4.0软件构建了其系统进化树。根据序列分析和系统进化关系分析的结果,将5头实蝇幼虫样品鉴定为番石榴实蝇(Bactrocera correctaBezzi),并将本研究获得的2条COI序列在GenBank中注册,GenBank的登录号为HM590450和HM590451。