基于ITS序列对青海不同地区中国沙棘遗传变异的分析

为研究青海省不同地区中国沙棘的遗传变异,本研究采集青海省不同地区野生的中国沙棘、西藏沙棘和肋果沙棘叶片作为实验材料,提取沙棘叶片的DNA,对其内转录间隔区(ITS)进行PCR扩增和扩增产物测序检测。对中国沙棘的ITS序列进行同源性和差异性分析,以西藏沙棘和肋果沙棘为外类群构建系统发育树。结果表明,不同地区中国沙棘ITS序列的同源性很高,均在99.5%以上。聚类分析结果显示中国沙棘、肋果沙棘和西藏沙棘各自分为一组,与形态学分类中归属于三个不同的种的分类结果一致。11个中国沙棘样品的ITS1区序列长度均为307 bp,5.8S区均为167 bp,ITS2区均为264 bp,3个区域分别存在3个、1个和5个变异位点。说明青海省不同地区中国沙棘的ITS区基因发生了不同程度的变异,本研究为进一步研究沙棘ITS基因提供了理论依据。     

氟吗啉对大豆疫霉菌的抑制作用

防治大豆疫病主要措施之一是使用甲霜灵,但由于其作用位点单一,病菌易对其产生抗性突变,为选出可替代甲霜灵的新型杀菌剂防治大豆疫霉根腐病,采用多位点杀菌剂氟吗啉,室内离体条件下测定了氟吗啉等3种杀菌剂对供试大豆疫霉菌各生长阶段的抑制作用。结果表明：氟吗啉对所测定的菌株4个发育阶段均有显著的抑制作用,氟吗啉对大豆疫霉菌菌丝生长EC50 值为0.1906μg/mL,EC95 值为0.4735 μg/mL;在0.5 μg/mL 浓度下对大豆疫霉菌孢子囊形成和孢子萌发抑制率达100%,低于达到相同抑制效果的金阿普隆和甲霜灵;说明该药剂在离体条件下能够有效抑制大豆疫霉菌的生长,效果好于金阿普隆和甲霜灵。     

基于ITS 2序列的15种淫羊藿属药用植物分类鉴定研究

为探讨内转录间隔区2(ITS2)序列及其二级结构在15种淫羊藿属(Epimedium)中的鉴定效率。利用试剂盒提取叶片总DNA,对ITS2序列进行PCR扩展与测序,计算K2P遗传距离,构建系统发育树,预测其二级结构。结果表明,ITS2序列在15种淫羊藿属植物中的通用性好,扩增成功率与测序成功率均为100%。从K2P种间和种内遗传距离看,ITS2在淫羊藿属中的种间变异较小,部分种类种内遗传距离大于种间遗传距离,鉴定成功率约为60%。采用比对法(Blast)分析,ITS2鉴定效率为75%。由ITS2系统进化树可知,淫羊藿属与鬼臼属各为一支,易于分开。除薄叶淫羊藿、竹山淫羊藿、巫山淫羊藿外,其余种类均能较好的进行聚类;除粗毛淫羊藿、保靖淫羊藿外其余均不能与NCBI下载到的序列进行较好聚类。研究表明,15种淫羊藿属植物ITS2序列在ML系统进化树中具较好的鉴别能力,但淫羊藿属各样本种内与种间遗传较为混乱,遗传距离复杂,ITS2二级结构形态结构亦较为相似,将各种间进行区分存在一定难度,不适合单独作为该属条形码使用。     

苎麻疫霉对几种杀菌剂的离体敏感性测定

在实验室条件下,研究了苎麻疫霉(PhytophthoraboehmeriaeSawada)对甲霜灵、地茂散、代森锰锌和杀毒矾的离体敏感性。结果显示,甲霜灵、地茂散、代森锰锌和杀毒矾对苎麻疫霉的菌丝生长抑制EC50平均值分别为0.0295mg/L、2.0283mg/L、25.1452mg/L和2882.8708mg/L,表明供试苎麻疫霉菌株对甲霜灵表现极为敏感,最低达0.0194mg/L;而对杀毒矾抗药性极强,最高达10158.7122mg/L,且各菌株间的最大差异达49.9倍。利用SPSS软件分析表明,各供试杀菌剂对苎麻疫霉菌株的质量浓度EC50值之间均不存在显著相关性,表明苎麻疫霉对上述杀菌剂的抗性或敏感性间不存在交互作用。     

辣椒疫霉对双炔酰菌胺抗药性生理生化机制研究

本试验对辣椒疫霉亲本菌株和抗性菌株的生理生化指标进行了测定,初步研究了辣椒疫霉对双炔酰菌胺产生抗性的机制。结果表明:辣椒疫霉亲本菌株和抗性菌株的菌丝生长受NaC l和葡萄糖影响较小,且NaC l和葡萄糖不同浓度处理后的所有菌株之间渗透压也均不存在显著性差异,故得知NaC l和葡萄糖均不能为疫霉菌提供营养和抑制其渗透。双炔酰菌胺低浓度处理能够使抗性菌株菌体内渗漏出较多的内含物,但随着处理时间的延长和浓度的提高内含物渗漏反而减少;亲本菌株和抗性菌株菌体内可溶性蛋白含量和β-1,3葡-聚糖酶的活力差异显著:亲本菌株菌体内的可溶性蛋白含量和β-1,3葡-聚糖酶的活力均高于抗性菌株。随着双炔酰菌胺处理时间的延长,亲本菌株和抗性菌株的可溶性蛋白含量和β-1,3葡-聚糖酶的活力都呈下降趋势。由此得出:辣椒疫霉内含物通过细胞膜外渗,致使药液在菌体内的积累减少,最终使到达作用靶标药剂的实际浓度下降;同时菌体的可溶性蛋白含量降低、β-1,3葡-聚糖酶的活力下降可能是对双炔酰菌胺产生抗性的原因。     

辣椒种子萌发对环境汞污染的敏感性研究

用不同浓度氯化汞溶液培养辣椒种子,研究辣椒种子萌发对环境汞污染的敏感性。结果表明,辣椒种子萌发过程对环境汞胁迫敏感,其敏感度顺序为胚轴伸长〉胚根伸长〉种子萌发率。当环境汞浓度为0.5μmol／L时,胚根伸长抑制率为10.1％,略低于胚轴伸长抑制率;随着环境汞浓度的增加,胚根伸长抑制率比胚轴伸长抑制率上升得快,汞浓度为50μmol／L时达到50.96％,而胚轴伸长抑制率只有38.5％。     

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。     

疫霉根腐病菌毒素对大豆不同组织中多酚氧化酶的影响

大豆疫霉根腐病是危害大豆生产的世界范围的毁灭性病害,毒素在其致病过程中起着重要的作用。本文对大豆疫霉根腐病菌毒素胁迫下抗感不同大豆品种根、茎、叶中多酚氧化酶(PPO)活性的变化进行了初步研究,结果表明:适宜浓度的毒素(稀释100倍,浓度为0.0897mg/mL)处理后,抗病品种根、茎和叶中PPO活性在病程的大部分阶段与对照相比都升高,并且根中的PPO比茎叶中的PPO反应更敏感;而感病品种在整个病程中虽然在某些阶段较对照有一定的提高,但幅度不大,在病程其他阶段PPO下降幅度远大于升高幅度。而浓度相对较高的毒素(稀释50倍,浓度为0.1794mg/mL)处理后抗感品种根、茎和叶中PPO活性的变化较稀释100倍浓度毒素幅度小。     

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。     

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.     

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.     

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.     

Epistasis in the resistance of pepper to phytophthora stem blight (Phytophthora capsici L.) and its significance in the prediction of double cross performances

Summary Epistatic effects are present in the resistance to Phytophthora capsici in peppers (Capsicum annuum). Genetic statistical analysis shows that additive by additive epistasis as well as higher order epistatic effects contribute to the total variation of resistance, significantly. Higher order epistatic effects caused bias in the prediction of the performance of double crosses, the difference between observed and predicted performances being significant.The relative importance of higher order epistasis with respect to additive by additive epistasis seems to be correlated with the aggressiveness of the isolate of Phytophthora capsici.     

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.     

蝉蜕诱导对球孢白僵菌生物学特性及其毒力影响的研究

利用改良蝉蜕诱导培养基对球孢白僵菌进行诱导,以无蝉蜕PDA培养基作为对照,目的是比较蝉蜕诱导后球孢白僵菌菌株的生长速率、产孢量、孢子萌发率等多个生物学指标,及二者对3龄亚洲玉米螟幼虫的毒力。经蝉蜕诱导的球孢白僵菌菌株的产孢量、孢子萌发率以及对玉米螟幼虫毒力都高于对照组,且差异显著,对玉米螟幼虫毒力处理组LT₅₀为4.73天,对照组需要6.97天。同时经蝉蜕诱导过的菌株开始产孢时间和胞外蛋白酶活性都优于对照组,胞外蛋白酶活性的变化范围是1.497×10^-2~3.538×10^-2 IU/mL,但经诱导菌株的生长速率低于处理组。鉴于球孢白僵菌菌株经蝉蜕诱导后,其产孢量和孢子萌发率增加,并且能够增强对玉米螟幼虫毒力,可利用其进行球孢白僵菌菌种保存和改良。

     

乌桕和无患子容器苗育苗密度与苗木生长和质量的关系

为提高乌桕与无患子的苗木品质,研究育苗密度对乌桕与无患子的苗木生长形态变化及苗木品质的影响。试验以当年生乌桕与无患子幼苗为试验材料,设不同育苗密度5个处理,测定分析不同育苗密度下乌桕与无患子幼苗的生长进程以及干物质积累、叶片数、主根长形态指标。结果表明:不同育苗密度下,乌桕与无患子的生长进程差异十分显著;且无患子和乌桕的苗高随着密度的变大逐渐增加,但是当苗高增加到一定程度时,苗高并未随之增加;乌桕、无患子的单株、茎生物量表现为随着育苗密度的变大而逐渐增加。且乌桕与无患子的合格苗所占的百分率随其苗木密度的变大而降低,与之呈负相关关系。综合苗高、地径、单株生物量、茎生物量、幼苗形态因子分化参数、合格苗率、单位面积合格苗数等指标,无患子以育苗密度为280~342株/m2、乌桕以育苗密度为337~412株/m2时,其生长形态和植株品质比较协调,植株苗木品级合格率较高。     

Genetics of Resistance to Phytophthora capsici in the Pepper Line 'SCM-334'

In order to establish a genetic model of the resistance to Phytophthora capsici in Capsicum annuum genotype ‘SCM-334’, two experiments were conducted which included ‘SCM-334’, as well as, the susceptible line ‘Morrón INIA 224’ and their Fl, F2, F3 and BC1 generations. We also tried to introgress a recessive gene, hypothetically responsible for resistance to P. capsici, from ‘SCM-334’ into ‘Morrón INIA 224’. Inoculations were made, when plants had 4—6 leaves, by irrigating the culture substrate with a zoospore suspension of P. capsici isolate ‘Bl’. The χ² test was applied to segregating generations to determine how many genes were involved in resistance. The hypothesis which explained the obtained segregations best, although not completely, was one that assumes 3 genes in ‘SCM-334’;, with at least 3 alleles in the heterozygous condition or 4 at any position, present in any genotype in order to be resistant. The influence of the isolate's aggressiveness and the inoculation method on the results are discussed.