基于转录组测序开发糜子SSR标记

【目的】 以6份不同地理来源的糜子资源为试验材料,基于前期转录组测序获得的1 000对SSR引物,选出200对进行多态性检测,以期构建一批可以准确评估糜子种质遗传差异的分子标记。【方法】 用Primer Premier 5.0软件设计引物,改良CTAB法提取DNA,PCR扩增DNA和聚丙烯酰胺凝胶电泳筛选引物多态性;用PowerMarker 3.25和PopGen 1.32计算遗传多样性参数。【结果】 200对引物中97对呈单态性,80对呈多态性。单碱基序列重复引物有20对,10对具多态性,其重复基元是A（50%）和T（50%）。二碱基序列重复引物有36对,15对具多态性,其碱基重复类型有7种（AG最多,TC、GC和GA次之,CA、TA和AC最少）。三碱基序列重复引物有144对,55对具多态性,其碱基重复类型有24种（GGC、GCG和GCC最多,GAA、GCT和CGC等次之,ACC、AGG、CAG、CGT、AAG、AAC、TCG、CGA、ATT、CAA和CCA最少）。就引物分辨率(Rp)值而言,1—3碱基序列重复引物分别为0.67—4.67（平均2.07）、1.33—4.33（平均2.73）和0.67—4.00（平均1.83）。基于Rp值分析SSR分布频次,发现80个标记分布在5个区间：0—1、1—2、2—3、3—4和4—5,分别包含17（21.25%）、36（45.00%）、11（13.75%）、14（17.50%）和2个（2.50%）。就$\overline{\text{Rp}}$值而言,1—3碱基序列重复引物分别为0.33—0.67（平均0.51）、0.40—0.78（平均0.59）和0.33—0.83（平均0.59）。单碱基序列重复标记共检测到22个等位变异,每个位点为2—3个（平均2.2000）,其中8个和2个位点分别具2和3个变异;二碱基序列重复标记共检测到38个等位变异,每个位点为2—3个（平均2.5333）,其中7个和8个位点分别具2和3个变异;三碱基重复标记共检测到136个等位变异,每个位点为2—3个（平均2.4727）,其中29个和26个位点分别具2和3个变异。就多态性信息含量而言,1—3碱基序列重复引物分别为0.3750—0.5355（平均0.4293）、0.2392—0.7438（平均0.4293）和0.2392—0.7438（平均0.3989）。就多样性指数而言,1—3碱基序列重复分别为0.6365—1.0776（平均0.7497）、0.5623—1.0986（平均0.8339）和0.5623—1.0889（平均0.8312）。【结论】 基于转录组测序结果,检测200对SSR引物的多态性,发现177对（88.5%）可以扩增出完整条带,其中80对具多态性,多态率为40%。     

茶树CsWRKYIIcs转录因子的克隆及功能分析

【目的】以‘陕茶1号’为材料,克隆CsWRKYIIcs转录因子并分析序列特征,了解其在不同组织和非生物胁迫下的表达模式以及转录活性,为深入研究茶树在逆境胁迫中的功能提供理论依据。【方法】根据茶树基因组数据库注释的WRKY序列设计特异性引物,采用RT-PCR技术从‘陕茶1号’中扩增CsWRKYIIcs的cDNA序列,用生物信息工具分析序列的特点,实时荧光定量PCR（qRT-PCR）研究基因的表达模式,用酵母试验验证其转录活性。【结果】获得9条CsWRKYIIcs的cDNA序列,开放阅读框长度分别为561、960、936、978、897、912、720、1 008和969 bp,分别编码186、319、311、325、298、303、239、335、322个氨基酸。除了CsWRKYIIc7缺少锌指序列外,其他CsWRKYIIcs均含有1个WRKY保守结构域和典型的C₂H₂型锌指结构。不同物种中的WRKYIIcs具有相似的保守基序,而茶树CsWRKYIIcs和拟南芥、葡萄等双子叶植物的WRKYIIcs氨基酸序列相似性更高。另外,CsWRKYIIcs启动子区域均预测到多个与非生物胁迫相关的顺式作用元件,意味着其可能参与非生物胁迫响应。qRT-PCR结果表明,9个CsWRKYIIcs在根和花中的表达量高于茎和叶,具有一定的特异性。同时,在干旱、ABA、高温和高盐胁迫下被不同程度的诱导表达,其中CsWRKYIIc1和CsWRKYIIc7的表达量变化显著,与推定的顺式作用元件结果相符。酵母试验表明9个茶树CsWRKYIIcs均具有转录激活活性。【结论】克隆获得9个茶树CsWRKYIIcs转录因子,它们参与了茶树对ABA、干旱、高温和高盐胁迫的响应,也可能发挥转录激活因子的调控作用。CsWRKYIIc1和CsWRKYIIc7可作为候选基因深入研究茶树的抗逆功能。     

土壤日晒在绿色植保中的应用与展望

土壤消毒主要用于土壤病虫草害防控。过去几十年,土壤主要采用化学药品熏蒸消毒,不合理使用容易造成环境污染。土壤蒸汽消毒、火焰消毒和热水浇灌等需要专门的仪器设备,且存在能耗较高、灭生性强、容易破坏土壤结构等缺点。目前,土壤日晒技术受到意大利、美国、以色列等70多个国家农业科学家的广泛关注,但在中国依然处于初级阶段。本文总结了国内外学者关于土壤日晒对农业病虫草害、土壤肥力、农作物产量等方面的影响与应用,分析了土壤日晒存在的局限性,并对土壤日晒在未来农业绿色防控上的应用前景进行了展望,以期为农产品无公害生产提供理论依据和实践参考。     

Effect of apple cultivars and storage periods on the virulence of Neofabraea spp.

Bull’s eye rot is a typical quiescent postharvest apple disease in major fruit-growing areas. The susceptibility of different apple cultivars to Neofabraea spp. (N. vagabunda and N. malicorticis) was assessed, with Granny Smith showing the most resistance and Cripps Pink the most susceptibility. To assess the factors involved in conidial germination, Neofabraea spp. were grown on crude protein extracts (CPEs) collected from apple fruits at different storage periods. Fungal germ tube growth rate and pathogenic enzyme (cellulase and xylanase) activity were assessed. Results showed that CPEs collected after 2 and 4 months of storage progressively stimulated conidial germination and germ tube elongation, while a lesser effect was observed from CPEs after 1 month of storage. Xylanase proved to be the main degrading enzyme secreted by all the isolates, while cellulase was produced only by N. vagabunda isolates. Overall, the isolate ID02 was the most virulent, based on more rapid germ tube elongation and greater activity of the lytic enzymes.     

水稻温敏型叶片白化转绿突变体tsa2的表型鉴定与基因定位

水稻温敏型叶色突变体是研究植物光合作用、叶绿体结构和功能以及温度影响叶绿体发育的理想材料。利用甲基磺酸乙酯(EMS)诱变籼型水稻(OryzasativaL.)三系保持系西农1B,从其后代中筛选到一个突变性状稳定遗传的温敏型叶片白化转绿突变体tsa2 (temperature-sensitive green-revertible albino 2)。与野生型相比, tsa2突变表型受温度影响, 22°C条件下萌发的野生型幼苗表型正常,而tsa2幼苗完全白化,且约40%白化苗死亡,存活白化苗的光合色素含量、光合速率均显著降低,成熟期主要农艺性状均显著变劣;在28°C下萌发的tsa2幼苗叶片呈浅绿色并伴有白条纹,其光合色素含量显著降低,光合速率及主要农艺性状差异较小; 32°C下萌发的tsa2幼苗叶片无明显差异。透射电镜观察显示,与野生型相比,tsa2在22°C下叶肉细胞中无叶绿体或存在异常发育叶绿体(尚未分化出基粒和基层),在28°C下部分叶肉细胞含少量发育完整的叶绿体,在32°C下叶肉细胞数量及形态均正常。实时荧光定量PCR(qRT-PCR)分析表明,与野生型相比,tsa2突变体中部分光合色素代谢途径基因、叶绿体发育相关基因及光合作用相关基因的表达水平呈不同程度变化。遗传分析表明, tsa2突变表型受一对隐性核基因控制, TSA2被定位于第5染色体SSR标记S5-57和S5-119之间,物理距离为718 kb。本研究为水稻遗传改良及研究温度影响叶绿体发育机制奠定了基础。     

Study on the Hepatoprotective Effect of Oxalis coriniculata L. and Related Mechanism by Regulating Oxidative Stress and TLR-2/NF-κB Signaling Pathway

[Objectives]This study aimed to explore the protective effect of Oxalis coriniculata L.on rats with acute liver injury induced by carbon tetrachloride(CCl4)and ...     

两种有机硅助剂对四氯虫酰胺防治菜青虫的增效作用

为明确两种有机硅助剂Silwet 408和Greenwet 7618对四氯虫酰胺防治菜青虫的增效作用,本文采用喷雾法测定四氯虫酰胺以推荐剂量、50%推荐用量分别与Silwet 408和Greenwet 7618的3 000倍稀释液混用对菜青虫的田间防效。结果表明,四氯虫酰胺以50%推荐用量分别与Silwet 408和Greenwet 7618混用,药后3 d对菜青虫的防效均与单独使用推荐剂量的防效无显著差异;药后7 d,50%推荐用量与Greenwet 7618混用对菜青虫的防效与推荐剂量的四氯虫酰胺仍无显著差异,而50%推荐用量与Silwet 408混用的防效显著降低;在药后14 d,所有处理对菜青虫无防治效果。因此,建议在生产上推广50%推荐用量的四氯虫酰胺与Greenwet 7618的3 000倍液混用防治菜青虫。     

Salvianolate attenuates oxidative stress injury of human endothelial EA.hy926 cells induced by hydrogen peroxide

AIM: To investigate the effect of salvianolate on oxidative damage induced by hydrogen peroxide in human endothelial EA.hy926 cells.METHODS: EA.hy926 cells were cultured in vitro and divided into the following groups:control group, damage group, and anti-damage groups (salvianolate+damage groups). The cell viability was measured by CCK-8 assay. The migration ability of the EA.hy926 cells was detected by Transwell assay. The content of nitric oxide (NO) in the culture supernatant of the EA.hy926 cells was examined. The levels of vascular endothelial growth factor (VEGF) were detected by ELISA. The apoptosis,mitochondrial membrane potential and intracellular superoxide anion content of the EA.hy926 cells were analyzed by flow cytometry. The protein levels of caspase-3, cleaved caspase-3, Bcl-2, Bax, NF-κB and p53 were determined by Western blot. RESULTS: Compared with damage group, the viability of EA.hy926 cells pretreated with salvianolate at different concentrations was significantly increased (P<0.05). The apoptotic rate was significantly decreased (P<0.05). Savianolate enhanced the migration ability of the cells. The levels of VEGF, NO and mitochondrial transmembrane potential were increased (P<0.05), and the intracellular ROS level was significantly decreased (P<0.05). The protein levels of NF-κB, p53, Bax and cleaved caspase-3 were significantly decreased, and the protein level of Bcl-2 was markedly increased(P<0.05). CONCLUSION: Savianolate reduces the damage of EA.hy926 cells by hydrogen peroxide exposure, and its mechanism may be related to the blocking of NF-κB signaling pathway.     

樱桃番茄抗青枯病砧木新品种海茄砧1 号的选育

海茄砧1 号是以野生茄子自交系HZ09-5 为母本，以从越南引进的茄子品种自交系HZ10-2 为父本育成的高抗青枯 病樱桃番茄砧木品种。植株生长势较强，第1 雌花节位约为第7 节，花为紫色，果实卵圆形，青熟果淡绿紫色或淡绿色，果 萼绿色，果长7～9 cm，果宽4～6 cm，单果质量为50～60 g，种子千粒重为3.2 g 左右，种皮浅黄色，高抗青枯病。海茄砧 1 号与樱桃番茄的嫁接亲和性好，共生性强，嫁接苗成活率高。嫁接后的樱桃番茄果萼开展，果面有光泽，可明显改善果实 VC、可溶性固形物和可滴定酸含量等品质，提高产量。适宜华南地区樱桃番茄的嫁接栽培。     

A rapid,simple, and sensitive immunoagglutination assay with silica nanoparticles for serotype identification of Pseudomonas aeruginosa

An agglutination test based on colored silica nanoparticles (colored SiNps) was established to detect serotypes of Pseudomonas aeruginosa. Monodisperse colored SiNps were used as agglutination test carriers. The colored SiNps were prepared through reverse microemulsion with reactive dyes, sensitized with 11 kinds of mono-specific antibodies against P. aeruginosa, and denoted as IgG-colored SiNps. Eleven kinds of IgG-colored SiNps were individually mixed with P. aeruginosa on a glass slide. Different serotypes of P. aeruginosa could be identified by agglutination test with evident agglutination. The P. aeruginosa could be detected in a range from 3.6 × 10⁵ to 3.6 × 10¹² cfu mL^?1. This new agglutination test was confirmed to be a specific, sensitive, fast, easy-to-perform, and cost-efficient tool for the routine diagnosis of P. aeruginosa.