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。     

中草药与苯扎溴铵联用治疗车轮虫病的案例分析

车轮虫病是鱼类重要的常见纤毛虫病。近年来,对传统化学药物耐药的车轮虫和小车轮虫越来越多。本文介绍了复方中草药联合苯扎溴铵治疗河鲀、黄颡鱼和草鱼车轮虫病的典型案例,以期为类似病例治疗提供参考。     

草原红牛Acot2基因克隆及其在各组织中的表达差异研究

本研究旨在探究草原红牛酰基辅酶A硫酯酶2（Acot2）的基因功能，并对其进行生物信息学分析，检测Acot2基因在草原红牛不同组织中的表达差异。根据GenBank中公布的牛Acot2基因序列（登录号：NM_001101938.1）设计引物，PCR扩增获得草原红牛Acot2基因的完整CDS并进行测序，利用分析软件进行序列同源性比对并构建系统进化树；获得对应的氨基酸序列并分析蛋白理化特性及蛋白亚细胞结构、亲疏水性和磷酸化位点，预测蛋白二级结构并构建蛋白质三级结构模型；利用实时荧光定量PCR方法检测Acot2基因在不同组织中的表达差异。结果显示，草原红牛Acot2基因CDS大小为1 395 bp，编码464个氨基酸，其核苷酸序列与亚洲水牛的同源性较高（98.3%），与猕猴和黑猩猩的同源性较低（80.5%和80.4%）。Acot2蛋白分子式为C₂₃₁₇H₃₆₀₆N₆₄₀O₆₂₈S₁₄，分子质量为50.924 ku，理论等电点为8.84。蛋白质不稳定指数为37.50，氨基酸残基多数为亲水性残基，总平均亲水性为-0.094。亚细胞定位分析表明，Acot2蛋白分布在内质网（30.4%）、线粒体（26.1%）、高尔基体（17.4%）、细胞质（17.4%）、液泡（4.3%）和细胞质（4.3%）中；磷酸化位点分析发现，Acot2蛋白存在20个磷酸化位点。二级结构主要形式有α-螺旋（21.8%）、β-转角（33.4%）、β-折叠（18.4%）和无规则卷曲（26.4%），三级结构预测结果与其相一致。实时荧光定量结果显示，Acot2基因在草原红牛胃中表达量最高，在肺脏中表达量极少。Acot2基因在生物进化过程中具有低保守性，其编码氨基酸组成的蛋白质结构稳定，属于水溶性蛋白，在线粒体和内质网中发挥作用，在草原红牛不同组织的表达量有明显差异。本研究结果为进一步探究Acot2基因对家畜脂代谢的影响和筛选草原红牛肉质候选基因提供资料。     

利用CRISPR/Cas9敲除葡萄VviPDS1基因的研究

选择葡萄八氢番茄红素脱氢酶(Phytoene desaturase)基因VviPDS1为靶标,利用CRISPR/Cas9系统构建基因敲除载体,瞬时转化葡萄叶片原生质体,检测到不同类型的突变。通过农杆菌介导转化‘无核白’葡萄胚性愈伤组织,筛选获得卡那霉素抗性植株71株。经PCR鉴定,其中53株为阳性植株,阳性率为74.64%。测序结果表明,共有20株在靶点发生不同类型的突变,编辑效率为37.74%;其中9株产生了双等位基因突变。对其进行氨基酸序列预测,在第202位氨基酸之后发生了不同程度的变异。利用CRISPR/Cas9系统敲除VviPDS1获得的突变体植株呈现整体矮化,其叶片出现不同程度白化。表明CRISPR/Cas9系统可以通过细胞中的瞬时或稳定表达进行基因编辑,可以实现在葡萄编辑植株中产生纯合敲除。     

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.     

Response of the soil macrofauna abundance and community structure to drought stress under agroforestry system in southeastern Qinghai-Tibet Plateau

ABSTRACT

Soil macrofauna is vital for soil functions and soil-mediated processes in all ecosystems. However, environmental perturbations, such as drought, that threaten both the abundance and function of soil macrofauna remain mostly unexplored, particularly in an agroforestry system. We investigated the effects of drought on soil macrofauna abundance and vertical distribution under three different planting systems including two intercropping systems, comprising Chinese prickly ash (Zanthoxylum bungeanum) intercropped with soybean (Glycine max) (Z-G) or bell pepper (Capsicum annuum) (Z-C), and one monoculture system, comprising only Z. bungeanum (Z). Soil samples were collected at depths of 0–10, 10–20, and 20–30 cm, and soil macrofauna and chemical properties were analyzed. Soil dryness negatively affected soil macrofauna in all planting systems. Drought reduced the total macrofauna density, biomass, genera richness, and Pielou’s evenness. Additionally, drought significantly decreased density and biomass of Drawida and Eisenia but had no effect on Carabid beetles. Soil macrofauna density was highest in the Z-G intercropping system and higher at 0–10 cm than at other soil depths. These results indicate that intercropping soybean rather than bell pepper increases the abundance and biomass of soil macrofauna, and drought remarkably impacts the response of soil macrofauna to planting systems.     

具有解析式位置正解的2T1R并联机构运动性能分析

求解具有解析式位置正解且部分运动解耦的并联机构,有利于后续的误差分析、动力学分析、运动轨迹规划与控制等。基于方位特征方程(POC)的并联机构设计理论与方法,设计了两种具有解析式位置正解且部分运动解耦的2T1R并联机构,并对这两种机构进行了方位特征、自由度及耦合度等主要拓扑性能分析;提出基于拓扑特征的运动学建模与求解方法,并据此求解了两种机构的解析式位置正解;基于导出的位置反解,分析了两种机构工作空间、奇异位形、动平台的速度与加速度变化规律。最后比较了两种机构的运动性能,选择了优选机型。为优选机型的动力学分析与样机研制提供了理论基础。