基于HVI测试的标准化操作规范

为了规范大容量棉花测试仪的使用、校准及维护,提高检测结果准确性.作者根据HVI应用手册和实际操作经验,总结了大容量棉花测试仪的标准化工作各项要求.主要包括样品管理、气电及空间要求、恒温室、标准化仪器、校准与核查、比对试验、测试人员、健康安全、消防安全等.供业内同仁参考.     

Semi-Synthesis,Cytotoxic Evaluation,and Structure—Activity Relationships of Brefeldin A Derivatives with Antileukemia Activity

Brefeldin A (1), a potent cytotoxic natural macrolactone, was produced by the marine fungus Penicillium sp. (HS-N-29) from the medicinal mangrove Acanthus ilicifolius. Series of its ester derivatives 2–16 were designed and semi-synthesized, and their structures were characterized by spectroscopic methods. Their cytotoxic activities were evaluated against human chronic myelogenous leukemia K562 cell line in vitro, and the preliminary structure–activity relationships revealed that the hydroxy group played an important role. Moreover, the monoester derivatives exhibited stronger cytotoxic activity than the diester derivatives. Among them, brefeldin A 7-O-2-chloro-4,5-difluorobenzoate (7) exhibited the strongest inhibitory effect on the proliferation of K562 cells with an IC₅₀ value of 0.84 µM. Further evaluations indicated that 7 induced cell cycle arrest, stimulated cell apoptosis, inhibited phosphorylation of BCR-ABL, and thereby inactivated its downstream AKT signaling pathway. The expression of downstream signaling molecules in the AKT pathway, including mTOR and p70S6K, was also attenuated after 7-treatment in a dose-dependent manner. Furthermore, molecular modeling of 7 docked into 1 binding site of an ARF1–GDP-GEF complex represented well-tolerance. Taken together, 7 had the potential to be served as an effective antileukemia agent or lead compound for further exploration.     

Comparative performance of the highly invasive Mimosa invisa and its less invasive subspecies under different nutrient and light conditions

Invasive plants may be more plastic than non‐invasive plants and maintain high fitness under various environmental conditions. Previous studies mainly focused on the comparisons between invasive and native plants, and comparisons between highly invasive and less invasive exotic species are still relatively rare, especially for comparisons at the subspecies level. This study examined the effects of nutrient addition and shading on the performance of the highly invasive Mimosa invisa and its less invasive subspecies M. invisa var. inermis under either isolated or competitive conditions. Nutrient addition increased biomass and plant height and decreased root‐to‐shoot ratio (R/S). Shading decreased biomass and R/S and increased plant height. Under isolated conditions, the two invaders did not differ in R/S, plant height and plasticity of these traits in response to nutrient addition or shading, and the two invaders also did not differ in biomass production under each of the nutrient and light treatments. When the two invaders competed with each other, M. invisa outcompeted M. invisa var. inermis under high soil nutrient conditions, and the two invaders did not differ in performance under other growth conditions. Thus, only considering competition may we find out the difference between highly invasive species and their closely related, less invasive subspecies. Management of M. invisa should focus on habitats with high soil nutrient availability, in which M. invisa is more likely to dominate.     

Evaluating diverse systems of tobacco genetic resistance to Phytophthora nicotianae in Yunnan,China

Black shank, caused by the soilborne pathogen Phytophthora nicotianae, is one of the most devastating diseases affecting tobacco production in China. The most effective strategy for reducing economic loss from this pathogen is development and use of resistant tobacco varieties. Multiple sources and systems of resistance have been developed in the Western Hemisphere; however, populations of P. nicotianae are variable around the world, including the predominance of different races. Different P. nicotianae isolates may react differently on tobacco plants with different systems of resistance, a possibility that could complicate the breeding of cultivars with resistance that is effective in different tobacco production regions worldwide. The objective of this research was to evaluate an array of tobacco germplasm possessing different systems of genetic resistance to black shank disease in tobacco-growing regions of Yunnan, China. Resistance types included simply inherited resistance mechanisms introgressed from wild Nicotiana relatives and polygenic partial resistance systems of N. tabacum origin. The loci of Wz exhibited high level resistance to black shank in the five diverse disease environments in Yunnan, China. K326 Php/−Wz/− genotype and Beinhart 1000 exhibited the greatest levels of resistance in both 2015 and 2016. Field observed results for 13 tobacco genotypes were highly correlated with those tested in growth chamber evaluation. These findings suggest that both Wz− and Beinhart 1000-mediated resistance have important commercial value in flue-cured tobacco breeding programmes in China. Cultivars developed for black shank resistance in China may also have utility in other tobacco-growing areas.     

云南省六个国家级自然保护区外来入侵草本植物与本地植物的关系

为明确不同自然保护区外来入侵草本植物及其与本地植物多样性之间的关系,以我国生物多样性丰富的云南省为研究对象,于2014年对云南省6个国家级自然保护区的82个1 m2小样方(78个100 m2大样方)中外来入侵草本植物和本地植物物种丰富度和密度进行调查,并分析外来入侵草本植物物种丰富度与本地草本植物物种丰富度、外来入侵草本植物密度与本地草本植物密度的关系及对不同自然保护区外来入侵草本植物物种丰富度的差异进行分析。结果显示,共调查发现22种外来入侵草本植物,其中纳板河流域国家级自然保护区种类最多,达14种,金平分水岭国家级自然保护区仅有2种;6个国家级自然保护区本地草本植物的物种丰富度与外来入侵草本植物的物种丰富度之间、本地草本植物密度与外来入侵草本植物密度之间均呈现正、负2种相关关系,其中哀牢山国家级自然保护区本地草本植物的物种丰富度与外来入侵草本植物物种丰富度呈显著负相关,其它5个国家级自然保护区的相关性均不显著。简单回归分析表明自然保护区外来入侵草本植物物种丰富度与本地草本植物物种丰富度、本地草本植物盖度、本地乔灌木物种丰富度呈显著负相关,多元回归分析结果表明乔灌木物种丰富度和经纬度是造成不同自然保护区外来入侵草本植物物种多样性差异的主要原因。     

棉铃虫脂肪酸结合蛋白的克隆与表达

为深入了解棉铃虫Helicoverpa armigera脂肪酸结合蛋白HaFABP1在参与细胞内脂肪酸代谢过程中的功能,基于酵母单杂交结果从幼虫中肠cDNA中扩增得到HaFABP1的开发阅读框,构建重组菌株BL21-pET28a-HaFABP1,进行融合蛋白His-HaFABP1的诱导和纯化,并利用实时荧光定量PCR检测棉铃虫不同时期和组织中HaFABP1的表达规律,最后检测2-十三烷酮处理下棉铃虫6龄幼虫中肠组织内HaFABP1的变化情况。结果显示,克隆获得的棉铃虫HaFABP1为405 bp,编码134个氨基酸,相对分子量和等电点分别为15.08 kD和5.54;在37℃下用0.5 mmol/L异丙基硫代半乳糖苷(IPTG)诱导重组菌株4 h后,得到约18.4 kD的可溶性蛋白His-HaFABP1,纯化后的融合蛋白条带单一且大小符合预期;HaFABP1在棉铃虫的脂肪体、中肠、体壁和头部中都有表达,在头部的表达量最低,在中肠中的表达量最高;HaFABP1在棉铃虫幼虫的每个发育时期都有表达,表达量总体呈现先降低后升高的趋势,4龄时最低而1龄时最高;10 mg/g 2-十三烷酮处理饲料饲喂棉铃虫6龄幼虫6 h和15 h后,中肠内HaFABP1的表达量与对照相比显著提高,且随着时间的延长逐渐增加。表明HaFABP1与棉铃虫的生长发育有关,并且有可能参与了昆虫的解毒过程。     

氯虫苯甲酰胺和氟虫腈对黏虫细胞色素P450基因表达的影响

为筛选出黏虫Mythimna separata参与杀虫剂解毒代谢的主效细胞色素P450基因,采用叶片浸渍法测定了用于处理黏虫3龄幼虫的亚致死浓度,通过构建转录组测序文库并结合数字基因表达(digital gene expression,DGE)对不同处理的黏虫进行测序,并运用实时荧光定量PCR(realtime quantitative polymerase chain reaction,RT-qPCR)技术验证12个P450基因的表达情况。结果表明,用于处理黏虫的氯虫苯甲酰胺和氟虫腈亚致死浓度LC_(10)分别为0.15、13.66 mg/L;对照样品、氟虫腈处理样品和氯虫苯甲酰胺处理样品分别获得59 521 504、64 838 148和41 722 990个原始序列数据,分别获得57 441 216、62 368 912和40 285 164个过滤后的序列数据;过滤后的序列长度分别为8.62、9.36和6.04 G;碱基错误率均为0.02%;Phred数值大于20、30的碱基占总碱基的百分比均高于90.59%;鸟嘌呤+胞嘧啶(guanine cytosine,GC)含量分别为47.16%、48.94%和47.55%,表明转录组测序质量较高;黏虫受氯虫苯甲酰胺胁迫后,29个P450基因表达量上调,27个P450基因表达量下调;黏虫受氟虫腈胁迫后,23个P450基因表达量上调,26个P450基因表达量下调;12个P450基因表达量的RT-qPCR技术检测结果与DGE测序文库显示的结果基本一致。     

天山野果林塞威士苹果和短距凤仙花叶斑病病原真菌的鉴定

为了解新疆天山野果林中塞威士苹果Malus sieversii与其林下伴生植物短距凤仙花Impatiens brachycentra两种植物叶斑病病原菌的多样性及同源性,采用组织分离法获得病原菌,基于rDNA-ITS序列构建系统发育树,并进行ITS序列BLAST同源性比对,对病原菌进行鉴定分类,并依据科赫氏法则测定致病性。结果显示,从新疆新源县天山野果林中的塞威士苹果及短距凤仙花病叶上共分离得到18株菌落形态各异的病原菌,分属于2属4种,绝大多数属于半知菌亚门。其中,链格孢属Alternaria sp.为塞威士苹果和短距凤仙花叶斑病病原真菌中的优势菌群。致病性测定结果显示,其中11株病原真菌对塞威士苹果具有致病性,7株病原真菌对短距凤仙花具有致病性;塞威士苹果所有的病原真菌对短距凤仙花均有致病性,而且短距凤仙花所有的病原真菌对塞威士苹果也有致病性。推测2种植物叶斑病可能由相同来源的病原真菌引发,短距凤仙花染病加剧了塞威士苹果叶斑病暴发,可能是野果林退化的重要原因。     

大气环流指数和地面气象要素对海南省橡胶树白粉病的影响

为探讨大气环流指数和地面气象要素对海南省橡胶树白粉病病情指数的影响,利用1962—2009年逐月大气环流指数和逐日地面气象要素资料,根据橡胶树白粉病病情指数等级,结合因子膨化、相关分析和多元回归等方法,筛选影响橡胶树白粉病病情指数的大气环流指数和地面气象要素,构建等级指标,建立预测模型,并对模型进行检验。结果显示:影响橡胶树白粉病病情指数的大气环流指数依次为北半球副高面积指数、北半球副高强度指数、西太平洋副高西伸脊点、北美区极涡面积指数、亚洲区极涡强度指数和印缅槽,显著影响时段为上年7月至当年4月;地面气象要素有平均最高气温、相对湿度、平均气温≥20℃的日数、小雨日数和平均风速,显著影响时段为上年10月至当年1月;基于大气环流指数、地面气象要素、大气环流与地面气象综合因子的橡胶树白粉病病情指数预测模型准确率分别为74.3%～78.8%、74.7%、94.6%～98.3%,表明基于大气环流和地面气象综合因子的模型准确率较高,可作为海南省橡胶树白粉病的中长期预测模型。     

中国茶树炭疽菌属病害研究进展及展望

茶树是一种重要的经济作物,广泛种植于热带和亚热带地区。随着茶产品经济价值的提高,茶叶生产安全备受关注。炭疽菌是引起茶树病害的病原菌之一,严重影响茶叶产量和茶产品品质,威胁我国茶产业的良性发展。利用抗病品种是防治茶树病害最为经济、有效的措施。目前,茶树炭疽菌种名使用混乱,茶树与炭疽菌的互作研究也较为薄弱,而简单依靠药剂防治病害也对茶叶质量安全造成一定的影响。本文梳理并明确了危害茶树的炭疽菌种类;总结了现有的茶树和炭疽菌互作的研究成果,提出识别免疫(effector triggered immunity,ETI)模式是茶树防御炭疽菌的主要机制;对茶树病理学存在的问题进行了归纳并提出解决办法,并展望了深入开展茶树抗病机理研究的思路,以期为制定茶园绿色防控技术及选育茶树抗病良种提供科学参考。