MOLECULAR CHARACTERIZATION OF AUTOPHAGY‐RELATED GENE 5 FROM Spodoptera exigua AND EXPRESSION ANALYSIS UNDER VARIOUS STRESS CONDITIONS

Autophagy is not only involved in development, but also has been proved to attend immune response against invading pathogens. Autophagy protein 5 (ATG5) is an important autophagic protein, which plays a crucial role in autophagosome elongation. Although ATG5 has been well studied in mammal, yeast, and Drosophila, little is known about ATG5 in lepidopteran insects. We cloned putative SeAtg5 gene from Spodoptera exigua larvae by the rapid amplification of cDNA ends method, and its characteristics and the influences of multiple exogenous factors on its expression levels were then investigated. The results showed that the putative S. exigua SeATG5 protein is highly homologous to other insect ATG5 proteins, which has a conserved Pfm domain and multiple phosphorylation sites. Next, fluorescence microscope observation showed that mCherry‐SeATG5 was distributed in both nucleus and cytoplasm of Spodoptera litura Sl‐HP cells and partially co‐localized with BmATG6‐GFP, but it almost has no significant co‐localization with GFP‐HaATG8. Then, the Western blot analysis demonstrated that GFP‐SeATG5 conjugated with ATG12. Moreover, real‐time PCR revealed that its expression levels significantly increased at the initiation of pupation and the stage of adult. In addition, the expression levels of SeAtg5 can be enhanced by the starvation, UV radiation, and infection of baculovirus and bacterium. However, the expression levels of SeAtg5 decreased at 24 h post treatments in all these treatments except in starvation. These results suggested that SeATG5 might be involved in response of S. exigua under various stress conditions.     

EXPRESSION AND EFFECTS OF MUTANT Bombyx mori ACETYLCHOLINESTRASE1 IN BmN CELLS

The main mechanism of toxicity of organophosphate (OP) and carbamate (CB) insecticides is their irreversible binding and inhibition of acetylcholinestrase (AChE), encoded by ace1 (acetylcholinestrase gene 1), leading to eventual death of insects. Mutations in AChE may significantly reduce insects susceptibility to these pesticides. Bombyx mori is an important beneficial insect, and no OP‐ or CB‐resistant strains have been generated. In this study, wild‐type ace1 (wace1) and mutant ace1 (mace1) were introduced into BmN cells, confirmed by screening and identification. The expression of wace1 and mace1 in the cells was confirmed by Western blot and their expression levels were about 21‐fold higher than the endogenous ace1 level. The activities of AChE in wace1 and mace1 transgenic cells were 10.6 and 20.2% higher compared to control cells, respectively. mace1 transgenic cells had higher remaining activity than wace1 transgenic cells under the treatment of physostigmine (a reversible cholinesterase inhibitor) and phoxim (an OP acaricide). The results showed that ace1 transgene can significantly improve ace1 expression, and ace1 mutation at a specific site can reduce the sensitivity to AChE inhibitors. Our study provides a new direction for the exploration of the relationship between AChE mutations and drug resistance.     

First Report of Black Leaf Spot Caused by Alternaria alternata on Ramie in China

In 2012 and 2013, black leaf spot disease was observed on ramie plants in Hunan and Hubei Provinces, China. In the field, the symptoms of this disease included dark green to black big spots on leaves, often resulting in upwardly curled leaf margins. The pathogen isolates were identified as Alternaria alternata (Fr.) Keissler on the basis of morphology and sequence similarity of 99–100% to the published data for internal transcribed spacer (ITS), and glyceraldehyde‐3‐phosphate dehydrogenase (gdp). To our knowledge, this is the first report of Alternaria leaf spot of ramie in China.     

高黎贡山南北段种子植物区系比较研究

为了解高黎贡山的种子植物资源,采用植物区系研究原理和方法,将高黎贡山南段与北段种子植物区系进行对比分析。结果表明,南段有192科878属2807种植物,种可分为15个分布类型9亚型14个变型;北段有172科778属2816种植物,种可分为15个分布类型9亚型20个变型。通过比较科属分化程度、数量构成及古热带成分比例等,南段区系具有较为古老和保守的特点,而北段植物区系具有较为年青和新生的特点。回归分析显示南北段区系地理成分相似度较高,而科属相似性却因分化在逐级降低。     

顽拗性三七种子后熟过程超微结构和抗氧化酶变化

该研究以3年生三七成熟种子为材料,通过对三七种子种胚切片观察、抗氧化酶活性测定及相关基因表达量的变化分析,从生理、形态及转录组3个层面了解顽拗性三七种子的内在机理.结果表明:三七种子后熟0~40 d,超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、抗坏血酸过氧化物酶(APX)和谷胱甘肽还原酶(GR)的活性先升高后降低,过氧化物酶(POD)的活性升高.后熟40 d时SOD、POD、CAT、APX相关差异表达基因的FPKM值分别为28、13、356、105,皆处于较高水平,此时观察到完整的细胞结构,种胚完成形态成熟,丙二醛(MDA)含量达到了最高值,说明三七种子内部抗氧化系统抵御氧化伤害最激烈,对水分胁迫造成的氧化伤害最为敏感.伴随后熟时间的延长,膜脂过氧化作用加剧造成细胞膜的降解,导致细胞功能丧失和畸形死亡,抗氧化系统酶活性降低不能有效抵御氧化伤害可能是导致顽拗性三七种子脱水敏感的重要原因之一.     

石灰岩山地优势种淡竹生物量分配的影响主因研究

该研究在江西省瑞昌市设置9个淡竹林样地,调查和测定了淡竹林密度、淡竹各构件的生物量和总生物量,以及土壤含水率、土层厚度、林下裸岩率、pH、电导率、全氮和全磷等7个土壤环境因子,并对竹林密度、土壤环境因子和淡竹生物量分配指标进行了相关分析和回归分析。结果表明：(1)密度与淡竹蔸比重相关系数r达0.66( P＝0.02<0.05),而与叶比重、枝比重、秆比重、鞭比重、根比重及根冠比均无显著相关关系;土壤环境因子与生物量分配指标有密切相关,环境主成分Z1与叶比重、秆比重及蔸比重均显著相关(P<0.05),Z2与鞭比重显著相关(P＝0.034<0.05)。(2)密度与土壤环境因子密切相关(P<0.05),控制土壤环境因子的偏相关分析显示密度与淡竹生物量分配不显著相关( P>0.05),而控制密度时,土壤环境因子与淡竹生物量分配仍有显著相关关系(P<0.05);逐步回归分析也验证了偏相关分析的结果,密度被排除出回归方程。分析认为,土壤含水率、土层厚度及土壤养分等环境因子是影响石灰岩山地优势种淡竹生物量分配的主因,密度对生物量分配的影响实为土壤环境因子的间接作用。该研究结果为石灰岩地区植被恢复提供了理论支撑。     

鸡肉粉完全替代鱼粉饲料中补充包被氨基酸对凡纳滨对虾生长、体成分及组织氨基酸含量的影响

为了探讨鸡肉粉完全替代鱼粉时饲料氨基酸的平衡性以及外源氨基酸的添加方式与凡纳滨对虾生长、体成分、血浆游离氨基酸及肌肉氨基酸含量的关系, 本试验采用26因子试验设计进行了为期56d的饲养试验。2个饲料蛋白质水平分别为40%和32%, 6个饲料处理分别为鱼粉组(对照组)、鸡肉粉组、鸡肉粉+晶体EAA组、鸡肉粉+晶体EAA+晶体NEAA组、鸡肉粉+包被EAA组、鸡肉粉+包被EAA+包被NEAA组, 配制12组饲料。将凡纳滨对虾(0.300.01) g随机分配到36个圆桶(150 L)中, 每桶30尾, 每3个桶为一个处理组, 饲喂一种饲料, 每天饱食投喂三次。在每一饲料蛋白质水平下, 无论是补充晶体氨基酸(CAA)组还是包被氨基酸组对虾的增重率均显著高于鸡肉粉组(P0.05), 且在32%蛋白质水平下, 包被EAA组对虾增重率达到了鱼粉组水平(P0.05); 补充晶体EAA+NEAA组对虾增重率与补充晶体EAA组无差异(P0.05), 但均显著低于补充包被氨基酸组(P0.05); 补充包被EAA组对虾增重率显著高于补充包被EAA+NEAA组(P0.05)。饲料系数的变化正好与增重率变化相反(P0.05)。饲喂高蛋白质水平饲料较之饲喂低蛋白质饲料明显提高对虾增重率、虾体蛋白含量(P0.05), 但降低虾体脂肪含量(P0.05)。包被氨基酸组凡纳滨对虾血浆游离氨基酸含量总体显著低于CAA组(P0.05)。除谷氨酸、甘氨酸以及脯氨酸外, 各组对虾肌肉氨基酸含量无显著差异(P0.05)。结果表明, 在32%饲料蛋白质水平下, 用鸡肉粉完全替代鱼粉时, 饲料中补充包被EAA可明显促进凡纳滨对虾的生长, 且达到了鱼粉组的饲喂效果。