3种含氯消毒剂对柑橘黄龙病菌室内防治效果研究

柑橘黄龙病是柑橘生产上最具毁灭性的病害，目前没有防治特效药也没有较好的抗性品种。本文以含氯消毒剂作为柑橘黄龙病菌防治的候选药剂开展药剂筛选试验。将染病接穗分别浸泡3种含氯消毒剂再嫁接到健康枳壳砧木上，利用定量PCR技术定量分析药剂处理前后黄龙病菌含量的变化，建立柑橘黄龙病菌药剂防治效果评价体系。试验结果表明，漂白粉（CH）不适合利用嫁接技术进行候选药剂筛选；二氯异氰尿酸钠（NaDCC）和三氯泡腾消毒片（TCCA）的抑菌效果随处理浓度的降低而减弱，在有效氯含量为5000mg/L时，病菌减退率最高，分别为99.1%和99.5%，相对防效最佳，分别达到了97.9%和98.8%，与对照药剂盐酸四环素(TET)的抑制效果相当，说明NaDCC和TCCA可以作为柑橘黄龙病化学防治的候选药剂。     

猪SENP1基因克隆、生物信息学分析与表达研究

试验旨在对猪SUMO特异性蛋白酶1(sentrin-specific protease 1,SENP1)基因CDS区进行克隆和生物信息学分析,并探讨SENP1基因在乙脑病毒(Japanese encephalitis virus,JEV)感染PK15细胞后的表达情况。根据GenBank中公布的猪SENP1基因预测序列(登录号:XM_013997974.2)设计特异性引物,通过RT-PCR和T/A克隆技术对猪SENP1基因CDS区序列进行克隆测序,并进行生物信息学分析,利用实时荧光定量PCR分析猪SENP1基因在JEV感染PK15细胞不同时间点(0、12、24、36、48 h)的表达情况。结果显示,猪SENP1基因CDS序列全长2 034 bp,共编码677个氨基酸。序列比对和系统进化树分析结果表明,猪SENP1蛋白序列和山羊、犬、人、牛的相似性分别为94.8%、94.2%、93.9%和93.8%,说明在这些物种中SENP1的保守性较强;猪与犬的SENP1分子亲缘关系较近。生物信息学分析结果显示,猪SENP1蛋白相对分子质量为76 825.76,等电点为8.53,体外半衰期为30 h,不稳定系数为53.59,总平均亲水指数为－0.622。SENP1蛋白不含信号肽序列,无跨膜结构域。二级结构分析结果显示,SENP1蛋白中α-螺旋、无规则卷曲、延伸链和β-转角分别占31.31%、46.68%、16.25%和5.76%。三级结构分析结果显示,猪SENP1蛋白中存在8个α-螺旋区和7个β-转角区。实时荧光定量PCR结果显示,猪SENP1基因在JEV感染的PK15细胞中呈现上调表达趋势,其中在感染后48 h SENP1基因表达量显著高于对照组(P<0.05)。本试验结果为后续深入研究SENP1基因功能提供了参考。     

绵羊FGF10基因克隆、序列分析及其在毛囊发育中的表达分析

本试验旨在获得中国美利奴羊成纤维细胞生长因子10（fibroblast growth factor 10，FGF10）基因的编码区（CDS）全长序列并进行生物信息学分析，随后对FGF10基因在中国美利奴羊毛囊发育过程中的表达特征进行分析，明确其在中国美利奴羊毛囊发育过程中的表达模式，为进一步研究FGF10 mRNA表达水平与中国美利奴羊毛囊生长发育的表达调控机制奠定理论基础。采用PCR扩增获得中国美利奴羊FGF10基因CDS，并克隆到zero PCR@^TM-Blunt进行测序验证；利用实时荧光定量PCR技术检测FGF10在中国美利奴羊毛囊发育过程中的表达差异。结果表明，绵羊FGF10基因CDS长度为696 bp（序列上传GenBank，获得登录号：MT872422），编码231个氨基酸，与牛和山羊的氨基酸序列同源性达100%，存在1个信号肽和1个跨膜结构域，其为分泌通路信号蛋白；实时荧光定量PCR分析表明，FGF10基因在中国美利奴羊毛囊发育过程中均表达，在毛囊发育第85天表达最高，显著高于其他毛囊发育时期（P<0.05）。本研究获得中国美利奴羊FGF10基因完整的编码区序列和毛囊发育过程中的表达特征，生物信息学分析发现，FGF10基因编码区序列具有物种间的保守性，同时FGF10在绵羊毛囊不同发育阶段的皮肤组织中表达，由此表明，FGF10基因可能在绵羊毛囊的生长发育过程中发挥重要的生物学作用。     

检测绵羊源爱知病毒D型荧光RT-PCR方法的建立和应用

绵羊源爱知病毒D型(ovine Aichivirus D)是在国内绵羊中新发现的病毒,本研究根据绵羊源Aichivirus D 3D基因序列设计检测引物,通过反应体系和条件优化,成功建立了检测绵羊源Aichivirus D的TB Green染料法荧光RT-PCR方法,该方法特异性和稳定性良好,灵敏度高。对2020年4月―2021年5月采集自四川6个场253份绵羊粪便样本(健康羊的133份,腹泻羊的120份)进行检测,结果该病毒的平均检出率为8.7％,场阳性率为66.7％。其中,腹泻粪便样本中绵羊源Aichivirus D阳性率(17.5%)显著高于非腹泻粪便样本中绵羊源Aichivirus D阳性率(0.75%,P<0.001)。表明绵羊源Aichivirus D可能是引起以上地区绵羊腹泻的病原。从绵羊源Aichivirus D的阳性样本中成功克隆出大小为1 422 bp的13个完整的绵羊源Aichivirus D 3D基因,其核苷酸相似性为99.4％～100.0％。遗传演化分析发现这13株绵羊源Aichivirus D 3D基因与本实验室前期研究上传的绵羊源Aichivirus D 3D基因共同聚为单独的一个大支。本研究为绵羊源Aichivirus D的分子检测提供了一种新的方法和基础流行病学数据。     

封育年限对典型草原大针茅无性系构件组成与生长的影响

研究不同封育年限对黄土高原大针茅(Stipa grandis)无性系构件结构组成和生长的影响,可为阐明无性系构件组成和资源分配提供一定基础。在黄土高原典型草原选取不同封育年限区(10和20年)和放牧地(对照)作为试验样地,采用整个无性系完整挖掘的方法进行大针茅无性系构件特征的研究。结果表明：1)与放牧相比,封育10年显著增加了大针茅无性系丛径、生殖枝高度和花序高度(P < 0.05),而封育20年则显著提高了大针茅无性系丛径、总枝数、生殖枝数、营养枝数、分蘖芽数、生殖枝高度、花序高度、地上总生物量、营养枝生物量和花序生物量(P < 0.05)。封育降低了大针茅生殖枝数、生殖枝高度、花序高度、地上总生物量、生殖枝生物量和花序生物量的变异系数。2)随封育年限增加,营养枝生物量比例明显提高,生殖枝生物量比例下降,但花序生物量比例基本不变。3)封育措施显著提高了营养株单株生物量(P < 0.05),但生殖株单株生物量和全体分枝单株生物量与放牧处理差异不显著(P > 0.05)。4)大针茅丛径与地上总生物量、生殖枝数、生殖枝生物量、花序生物量呈极显著正相关关系(P < 0.001),与营养枝生物量呈极显著正相关关系(P < 0.01),与总枝数呈显著正相关关系(P < 0.05)。而地上总生物量与生殖枝数、营养枝数、总枝数、营养枝生物量呈极显著正相关关系(P < 0.001),与分蘖芽数、生殖枝生物量和花序生物量呈极显著正相关关系(P < 0.01)。分蘖芽数与营养枝数、总枝数、营养枝生物量呈极显著正相关关系(P < 0.001),与地上总生物量呈极显著正相关关系(P < 0.01)。综上所述,短期封育提高了大针茅的生殖分配,使大针茅迅速成为群落优势种,而长期封育使大针茅更依靠无性繁殖进行种群更新。     

T1 locus in cotton is the candidate gene affecting lint percentage, fiber quality and spiny bollworm (Earias spp.) resistance

A genetic linkage map of chromosome 6 was constructed by using 270 recombinant inbred lines originated from an upland cotton cross (Yumian 1 × T586) F₂ population. The genetic map included one morphological (T₁) and 18 SSR loci, covering 96.2 cM with an average distance of 5.34 cM between two markers. Based on composite interval mapping (CIM), QTL(s) affecting lint percentage, fiber length, fiber length uniformity, fiber strength and spiny bollworm resistance (Earias spp.) were identified in the t₁ locus region on chromosome 6. The allele(s) originating from T586 of QTLs controlling lint percentage increased the trait phenotypic value while the alleles originating from Yumian 1 of QTLs affecting fiber length, fiber length uniformity, fiber strength and spiny bollworm resistance increased the trait phenotypic value.     

Genetics of Resistance in Vicia sativa L. to Orobanche crenata Forsk.

The genetics of resistance of common vetch (Vicia sativa L.) to broomrape (Orobanche crenata Forsk.) was studied for two years by using the P₁, P₂, F₁, BC¹, BC₂, F₂ F₃, and F₄ generations obtained from crosses between resistant and susceptible lines. Resistant lines were selected by screening a world collection m a naturally infested plot. Resistance was tested both under field and greenhouse conditions. The best index to measure resistance was the number of emerged broomrapes per host plant. The results fit the additive-dominance model. The main component of the variation was additivity; dominance and interaction effects seemed to depend on the environment. When dominance is expressed, a low number is dominant over a high number of broomrapes per host plant.     

Widening the gene pool of cultivated lentils through introgression of alien chromatin from wild Lens subspecies

Wild Lens species/subspecies are a potential source for increasing genetic diversity in cultivated lentil. Four intraspecific crosses were attempted between cultivated and wild lentils. Viable hybrids were produced between L. culinaris ssp. culinaris × L. culinaris ssp. orientalis and L. culinaris ssp. culinaris × L. culinaris ssp. odomensis. Normal meiosis and pollen fertility were observed in the first set of crosses, whereas chromosomal abnormalities and reduced pollen fertility were observed in the second set of crosses. These crosses were also studied for some quantitative traits. The range, mean and coefficient of variation were calculated in parents, F₁, F₂ and BC₁ generations to determine the extent of variability generated in the cultivated lentil through introgression of genes from wild lentil. The cultivated lentil × L. culinaris ssp. orientalis crosses showed substantially higher variability for all the traits than crosses involving cultivated lentil ×L. culinaris ssp. odomensis. The results of the present study indicated that these wild subspecies can be exploited for breeding purposes and their variation can easily be utilized to widen the genetic base of the cultivated lentil.     

QTL analysis of seed dormancy in rice (Oryza sativa L.)

Effective cumulative temperature (ECT) after heading would be a more reasonable parameter for seed sampling of pre-harvest sprouting/seed dormancy (SD) tests in segregating populations than the days after flowering. SD is an important agronomic trait associated with grain yielding, eating quality and seed quality. To identify genomic regions affecting SD at different grain-filling temperatures, and to further examine the association between SD and ECT during grain-filling, 127 double haploid (DH) lines derived from a cross between ZYQ8 (indica)/JX17 (japonica) by anther culture were analyzed. The quantitative trait loci (QTLs) and their digenic epistasis for SD were identified using a molecular linkage map of this population. A total of four putative QTLs for SD (qSD-3, qSD-5, 6 and 11) were detected on chromosomes 3, 5, 6 and 11, together explaining 41.4% of the phenotypic variation. Nine pairs of digenic epistatic loci were associated with SD on all but chromosome 9, and their contributions to phenotypic variation varied from 2.87%–8.73%. The SD QTL on chromosome 3 was identical to the QTLs found in other mapping populations with different genetic backgrounds, which could be a desirable candidate for gene cloning and marker-assisted selection in rice breeding.     

Identification of quantitative trait loci underlying milling quality of rice (Oryza sativa) grains

Milling quality of rice grains is important to both producers and consumers. In this study, quantitative trait loci (QTLs) controlling brown rice rate (BR), milled rice recovery (MR) and head rice recovery (HR) were analysed by composite interval mapping over 2 years using 98 backcross inbred lines (BILs). A total of 12 QTLs for the three traits were detected, of which five were for BR, four for MR and three for HR. The proportion of phenotypic variation explained by individual QTLs ranged from 7.5 to 19.9%, and additive effects contributed by a single QTL accounted for 0.46 to 2.34% of the variation. QTL‐by‐environment interactions were observed by comparing QTL mapping of the same population grown in two consecutive years. Three of five QTLs for BR and two of four QTLs for MR were detected in 2 years, and all three QTLs for HR were detected in 1 year only. BR was significantly correlated with MR, and all four QTLs of MR were located in the same regions as those of BR. This indicated that QTLs for highly correlated traits could often be detected in the same interval.