Progresses in the Mechanism of Resistance to Fusarium Wilt in Cucumber （Cucumis sativus L.）

Fusarium wilt caused by Fusarium oxysporum f.sp. cucumerinum （Owen） is one of the most devastating diseases in cucumber production worldwide. Recent progresses in the mechanism of resistance to Fusarium wilt in cucumber were reviewed in this paper, including pathogenic mechanism of Fusarium oxysporum, the resistance mechanism of cucumber, the heredity of resistance, and the location of resistance genes. Following works should be the location and cloning of resistance genes with molecular biologic methods.     

Characterization of RAPD Markers, and the RFLP Marker Linked to Powdery Mildew Resistance Gene Derived from Different Accessions of H. villosa

The analysis was carried out on performance of the resistance gene from Haynaldia villosa accession of the former Soviet Union to different isolates ofBluemerie graminis. Polymorphisms were revealed between 6D/6V substitution line Pm930640 and its pedigree parents using five RAPD markers of OPAN031700, OPAI01700, OPAL03750, OPAD07480 and OPAG 15580 screened out from 120 random 10-mers primers. Three RAPD markers of OPAN03, OPAI01 and OPAL03 were linked with the resistance gene by analysis of F2 population of ChancellorxPm930640. Analysis of 29 wheat lines including part of lines conferring the known genes from Pro1 to Pro20 respectively, lines conferring resistance gene from two H. villosa accessions and the related wheat parents, were analyzed and the results showed that these markers not only linked to the gene resistant to powdery mildew from H. villosa, but also detected different genetic backgrounds. OPAL03750 can be used as the marker to distinguish the different resistant lines from two H. villosa accessions because it was only observed in the materials from H. villosa of the former Soviet Union. RFLP analysis also showed the polymorphisms between two H. villosa accessions and their derived resistant lines.     

Evaluation of Some Physical Properties of Cucumber （Cucumis sativus L.）

Cucumber is one of the important vegetables in many countries. Information about physical properties of cucumber is necessary for designing, grading, sorting and processing operations. In this study, some physical properties such as diameter, length, height, and weight of three varieties of cucumber （Green Gold, Dharwad, and Super Dominus） were measured. The results revealed that there was no significant difference （P〈0.01） in the density values of the studied varieties, but regarding other physical properties there were a significant difference （P〈0.01） among them. The average density for Green Gold, Dharwad and Super Dominus were 0.98, 0.95 and 0.94 g/cm^3, respectively. Green Gold cucumber had the highest diameter, volume, weight, flesh diameter, geometric mean diameter and sphericity. Dharwad and Super Dominus varieties with 15.49 cm length and 4.61 length to diameter ratio had the highest length and length to diameter ratio, respectively. Smallest skin thickness （1.48 cm） was for Super Dominus. Surface area of Green Gold, Dharwad and Super Dominus were 192.29, 192.4 and 131.2 cm^2. As well as there was a high and positive correlation between weight and volume in the studied varieties. There was a non-significant and low correlation （0.56） between diameter and length in Super Dominus. There was also a high and positive correlation between diameter and geometric mean diameter in all the varieties.     

Identiifcation and Molecular Mapping of the RsDmR Locus Conferring Resistance to Downy Mildew at Seedling Stage in Radish （Raphanus sativus L.）

Downy mildew （DM）, caused by the fungus Peronospora parasitica, is a destructive disease of radish （Raphanus sativus L.） worldwide. Host resistance has been considered as an attractive and environmentally friendly approach to control the disease. However, the genetic mechanisms of resistance in radish to the pathogen remain unknown. To determine the inheritance of resistance to DM, F1, F2 and BC1F1 populations derived from reciprocal crosses between a resistant line NAU-dhp08 and a susceptible line NAU-qtbjq-06 were evaluated for their responses to DM at seedling stage. All F1 hybrid plants showed high resistance to DM and maternal effect was not detected. The segregation for resistant to susceptible individuals statistically iftted a 3：1 ratio in two F2 populations （F2（SR） and F2（RS））, and 1：1 ratio in two BC1F1 populations, indicating that resistance to DM at seedling stage in radish was controlled by a single dominant locus designated as RsDmR. A total of 1 972 primer pairs （1 036 SRAP, 628 RAPD, 126 RGA, 110 EST-SSR and 72 ISSR） were screened, and 36 were polymorphic between the resistant and susceptible bulks, and consequently used for genotyping individuals in the F2 population. Three markers （Em9/ga24370, NAUISSR826700 and Me7/em10400） linked to the RsDmR locus within a 10.0 cM distance were identiifed using bulked segregant analysis （BSA）. The SRAP marker Em9/ga24370 was the most tightly linked one with a distance of 2.3 cM to RsDmR. These markers tightly linked to the RsDmR locus would facilitate marker-assisted selection and resistance gene pyramiding in radish breeding programs.     

Bioinformatics Analysis on Regulating Axillary Branch Gene of Cucumber（Cucumis sativus L.）

The regulating axillary branch gene was cloned and named as CsCCD7.Using a series bioinformatic computer softwares,database and online programes,CsCCD7 nucleotide sequence and CsCCD7 amino acid sequence were analyzed and CsCCD7 function was predicted.The results showed that CsCCD7 cDNA full length sequence was 2 136 bp,and included a 1 665 bp ORF which encoded a 554 AA protein;there were 32 kinds of cis-acting regulating element in 2 136 bp cDNA sequence;CsCCD7 was an unstable protein(the unstable coefficie...     

Cytological Analyses on Development of Male and Female Gametophytes in an Interspecific Hybrid F1 from Cucumis hystrix Chakr. x Cucumis sativus L.

An interspecific hybrid F1 of Cucumis hystrix Chakr. x Cucumis sativus L. （NC4406） was used to establish the developmental sequence and to characterize the male and female gametophytes at cytological level for further understanding of the phylogenic relationship and the mechanism of fertility or sterility in the interspecific hybrid F1 The development of male and female gametophytes was studied through meiotic analysis and paraffin section observation technique, respectively. Meanwhile, the fertility level was assessed through hybrid F1 backcrossing to cultivated cucumber 4406. Variable chromosome configurations were observed in the pollen mother cells （PMCs） of hybrid F1 at metaphase Ⅰ , e.g., univalents, bivalents, trivalents, quadravalents, etc. At anaphase Ⅰ and Ⅱ, chromosome lagging and bridges were frequently observed as well, which led to the formation of polyads and only a partial number of microspores could develop into fertile pollen grains （about 23.3%）. Observations of the paraffin sections showed numerous degenerated and abnormal embryo sacs during the development of female gametophytes, and only 40% of the female gametophytes could develop into normal eight-nuclear megaspore. On an average, 22.8 and 6.3 seeds per fruit could be obtained from the reciprocal backcross. The interspecific hybrid F1 of C. hystrix x NC4406 was partially fertile; however, the meiotic behaviors of hybrid F1 showed a high level of intergenomic recombination between C. hystrix and C. sativus chromosomes, which indicated that it plays an important role for introgression of useful traits from C. hystrix into C. sativus.     

Germplasm Evaluation,Inheritance,Molecular Marker and Mechanism of Resistance to Frog—eye Leaf Spot

Soybean frog-eye leaf spot(FLS)has been a major disease and production constralnt in the Northeast of China.The paper reported methods for identifying and evaluating resistant germplasm to the disease,at sometime listed several resistant cultivers or lines which were resistant to both dominant races(1,7race)and the mixture of over 8 races.It indicated that the resistance to race 1 or 7 were due to two different single dominant genes,respectively.And in he fied,there were more races,the resistance showed quantitative character.It was proposed the LTP,which was made according to the sizeand number of lesions could be used as an important index in selecting.Using bulked segregants analyxsis(BSA),the genetic distance between OPSO3 620 and resistant gene to race 7 wer identified 8.7cm.he results showed that the resistant cultivars had less number of stoma,closer arrangement of poliseade tissue and high cotent of wax.     

黄瓜白粉病抗性基因的QTL定位

 【目的】对黄瓜高抗白粉病材料K8进行研究,明确其抗性遗传规律,并完成抗性基因的QTL定位分析,为探索抗病机理和分子标记辅助选择（MAS）育种提供理论依据。【方法】利用黄瓜白粉病致病菌Podosphaera xanthii (syn. Sphaerotheca fuliginea）对K8×K18（感白粉病）杂交后代F2:3家系人工接种鉴定,进行抗白粉病遗传分析。以完成抗病性鉴定的F2和F2:3家系组成的抗感分离群体为研究对象,应用BSA法和2360对黄瓜SSR引物进行SSR分析,采用JoinMap 4.0作图软件和MapInspect软件构建连锁群并完成连锁群与染色体的对应。利用MapQTL4.0软件对白粉病抗性基因进行QTL定位分析。【结果】试材K8所含有的黄瓜白粉病抗性基因符合数量性状遗传的特点。本研究共检测到4个白粉病抗性基因的QTL位点pm5.1、pm5.2、pm5.3和pm6.1,其中,pm5.1、pm5.2、pm5.3在两年中被重复检出,pm5.2位点的贡献率最大,在其所在区域预测到了4个NBS类抗病基因。pm6.1位于黄瓜Chr.6上,是个微效的QTL位点。【结论】位于Chr.5上的pm5.2是黄瓜白粉病抗性基因的主效QTL位点,该抗性基因可能属于NBS类抗病基因。本研究结果为抗性基因主效QTL的精细定位和克隆及MAS抗病育种奠定了良好基础。     

黄瓜耐冷性遗传分析与连锁标记筛选

黄瓜(Cucumis sativusL.)是典型的冷敏型植物。对黄瓜来说,冷害是生产上制约其丰产、优质的主要逆境因素之一。为了掌握黄瓜耐冷性遗传规律,加快黄瓜耐冷品种的选育,本研究选取黄瓜耐冷型品系0839和低温敏感型品系B52为亲本杂交得到F1和F2,进行苗期低温鉴定和遗传分析。供试亲本的耐冷性主要受一对显性单基因控制。结合BSA(群体分离分析)和SSR分子标记,获得了与黄瓜耐冷性主效基因连锁的SSR标记。通过F2群体分析,鉴定出与耐冷性基因连锁的分子标记SSR07248,该标记与耐冷性基因间的遗传距离为32.6cM。     

黄瓜CC-NBS-LRR家族基因鉴定及在霜霉病和白粉病胁迫下的表达分析

【目的】对黄瓜全基因组的CC-NBS-LRR（CNL）基因家族进行生物信息学和表达模式分析,为深入研究CNL基因家族在黄瓜生长、发育和病害胁迫响应中的功能提供参考。【方法】以拟南芥CNL为参考序列,利用本地Perl语言和Pfam等软件检索黄瓜‘9930’基因组并确定黄瓜CNL基因家族成员。通过ExPASy、GSDS2.0、MEGA、MEME、Tbtools、Mev等工具对黄瓜CNL家族基因进行生物信息学分析。根据转录组数据库、霜霉病菌（Pseudoperonospora cubensis）、白粉病菌（Podosphaera xanthii）接种处理和实时荧光定量PCR技术分析该类基因的表达模式。【结果】从黄瓜全基因组中鉴定得到17个CsCNL,这些基因分布在除1号染色体外的6条染色体上,其编码蛋白与其他植物CNL结构相似,均含有CC、NBS和LRR保守结构域,蛋白质大小介于197—1 148 aa,分子量在22.6—131.3 kD,等电点在5.71—8.38。共线性分析表明其中不存在片段重复和串联重复基因,多物种系统进化关系表明,CsCNL基因家族成员在葫芦科植物之间具有较高的结构和功能相似性。CsCNL启动子中存在多种与抗病相关的顺式作用元件。CsCNL表达具有组织特异性。在接种霜霉病菌2 d和5 d后,Csa3G815400、Csa3G822360和Csa7G420890在抗性品种中均显著上调表达,Csa4G015850在抗性品种中下调表达,在敏感品种中显著性表现不一;在接种白粉病菌2 d和5 d后,Csa2G008000和Csa3G684170在敏感品种中显著上调表达,在抗性品种中显著下调表达;Csa4G016360、Csa7G420890和Csa7G425940在抗性品种中显著上调表达,在敏感品种中表达量无变化或显著下调。【结论】CsCNL家族成员具有组织表达特异性并且多数基因能够响应霜霉病菌和白粉病菌的胁迫。推测Csa3G815400、Csa3G822360和Csa7G420890表达量增加,Csa4G015850表达量减少可诱发抗病黄瓜品种的霜霉病抗病反应;Csa4G016360、Csa7G420890和Csa7G425940表达量增加,Csa2G008000和Csa3G684170表达量减少可诱发抗病黄瓜品种的白粉病抗病反应;而Csa7G420890表达量的增加能同时诱发抗病黄瓜品种的霜霉病和白粉病的抗病反应。     

外源BR诱导黄瓜(Cucumis sativus L.)幼苗的抗盐性

【目的】明确外源油菜素甾醇(Brassinosteroid,BR)对黄瓜(CucumissativusL.)幼苗抗盐性的诱导作用。【方法】采用根际注射结合叶面喷施外源BR(0、0.005、0.01、0.05、0.1、0.2mg·L-1)的方法,比较分析了盐胁迫下幼苗植株盐害指数、抗氧化酶活性、渗透调节物质含量等生理指标。【结果】外源BR能够明显改善盐胁迫下黄瓜幼苗植株的生长发育状况,降低盐害指数,最高幅度达40.2个百分点(P﹤0.01),极显著地提高叶片细胞游离脯氨酸(Pro)和可溶性糖的含量、超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)等抗氧化酶活性(P﹤0.01),进而保护细胞膜的稳定性。【结论】外源BR可以有效诱导黄瓜幼苗的抗盐性,并且最佳浓度范围是0.01～0.05mg·L-1。     

无土栽培黄瓜品种比较试验

对在土壤栽培中表现较好的三个品种“西杂”、“828”和“长春密刺”进行了无土栽培比较试验。观察测定了各品种的生长势、生育期、果实性状、早期产量和总产量。综合分析认为:黄瓜杂一代“828”较为适合无土栽培。     

黄瓜基因组DNA提取及AFLP体系优化研究

［目的］提取黄瓜基因组DNA,并对AFLP体系进行优化。［方法］以黄瓜嫩叶为材料,利用改进的CTAB法提取高质量的黄瓜叶片总DNA,通过优化酶切连接、预扩增、选择性扩增等试验条件建立适合黄瓜的AFLP银染体系,得到清晰的黄瓜AFLP指纹图谱。［结果］DNA 模板的质量影响酶切以及后续的连接扩增反应,改良的CTAB 提取法可用于黄瓜AFLP 分析,形成清晰的AFLP 指纹。优化的酶切连接体系为：以37 ℃酶切连接12 h为宜,酶切连接的基因组DNA用量为200 ng,预扩增时Taq酶用量为0.5 U/20 μl体系,预扩增产物稀释40倍作为选择性扩增的模板。在此优化的体系下引物E41M47扩增出清晰的条带。［结论］为黄瓜品种的分子标记和黄瓜品种间亲缘关系等研究奠定了基础。     

两个黄瓜品种对白粉病的抗性遗传分析

以美国的黄瓜抗病自交系WIS2757与津研2号自交系及相关杂交后代为材料,采用喷雾接种法,在空调温室内进行了黄瓜抗白粉病(Sphaerothecafuliginea)遗传规律的研究。结果表明,WIS2757对白粉病的抗性基因模式为RRssii,即抗性由1个隐性主效基因s,1个显性加强基因R和1个隐性基因i决定。津研2号与WIS2757抗性的差异主要在于R基因的差异,即津研2号的基因型模式为rrssii,抗性由1个隐性主效基因s和1个隐性基因i决定。感病亲本的基因组成为RRSSII。     

农杆菌介导的MADS-box基因转化黄瓜初步研究

试验通过叶盘转化法,利用根癌农杆菌介导,将从黄瓜中克隆到的MADS-box同源基因—CSMADS06基因,转化到烟草和2种不同品系的黄瓜S06和S52中,经过除草剂筛选获得14株转化烟草和28株转化黄瓜。PCR检测和GFP荧光检测证实,外源的CSMADS06基因已转入这些植株中。阳性率分别为2.36%(烟草),4.1%和2.9%(黄瓜)。抗性植株已开花结果并获得子一代植株。     

黄瓜白粉病抗性基因定位及候选基因分析

【目的】通过对黄瓜白粉病抗性基因的精细定位,明确抗性基因的候选区段,为进一步克隆基因及解析基因功能奠定基础。【方法】本研究选用高抗白粉病资源(自交系‘74’)和感白粉病资源(自交系‘80’)配制杂交组合,构建F_1、F_2群体,利用单囊壳白粉菌鉴定亲本及群体白粉病抗性并进行遗传分析;采用BSA-seq的方法对黄瓜白粉病抗性基因进行初步定位,在此基础上在候选区段开发分子标记,进一步精细定位白粉病抗性基因。【结果】分离群体单株表型鉴定结果表明,自交系74的抗性是由不完全隐性基因控制的。BSA-seq技术初步将抗性基因定位于5号染色体15—25 Mb位置,命名为PM 74。最终,利用分子标记将抗性基因定位于SSR15321和SSR07531之间,遗传距离为3.06 c M,物理距离为238 444 bp,可解释41.95%的表型变异。生物信息学分析表明在候选区段内包含有17个候选基因,其中Cucsa.275630为TIR-NBS-LRR类基因。【结论】本研究将黄瓜白粉病抗性基因精细定位到约238 kb的候选区段内,区段内包含有一个TIR-NBS-LRR基因,该基因将是今后研究的重点候选基因。     

黄瓜的分子标记和连锁图谱研究进展

黄瓜分子标记和遗传连锁图谱的研究工作相对番茄、小麦等作物比较落后,至今开展的分子标记研究多围绕黄瓜遗传关系分析进行,现有的几张遗传连锁图谱基本是由美国Staub研究小组完成,已被定位在图谱上的同工酶、RAPD、RFLP、SSR、AFLP等标记总数达到300多个,各标记间平均距离达到2.1cM,被整合在一起的遗传基因不足10个(F、B、de、ll、dm)。