白屈菜CmFAD2基因的克隆及植物转化载体构建

通过对白屈菜低温应答过程的转录组分析发现膜脂不饱和化相关基因的表达在一定过程中发生变化,脂肪酸去饱和酶基因FAD2在随温度的变化趋势为正"V"型,且表达量变化显著。利用NCBI等在线软件对序列进行相关生物学信息分析,并对白屈菜FAD家族成员FAD2基因的完整开放阅读框(ORF)进行克隆,并命名为CmFAD2。选用克隆载体pMD-19-T,转化大肠杆菌DH5α,测序验证序列正确性及完整性。将目的基因与植物表达载体pRI-201-AN连接构建重组DNA pRI-201-AN-Cm FAD2,电击法转化农杆菌LBA4404,利用菌液PCR法验证成功。该基因可作为药用植物抗寒品种创制的候选基因。     

Leaf carbohydrates assimilation and metabolism affect seed yield of rapeseed with different waterlogging tolerance under the interactive effects of nitrogen and waterlogging

Waterlogging is a main factor causing rapeseed (Brassica napus L.) yield loss, and reasonable nitrogen (N) applications can compensate for this loss. To investigate the effects of N rates on seed yield of waterlogged rapeseed, the waterlogging-tolerant rapeseed variety ZS 9 and sensitive variety GH01 were waterlogged for 0 and 10 days with five leaves at the seedling stage under four N rates (0, 90, 180 and 270 kg/ha). Waterlogging significantly decreased seed yield, while N application can alleviate the yield loss. The yield decrease rate of waterlogged GH01 was greater than that of ZS 9 under the same N rate. During the seedling and bolting stage, the leaf photosynthetic rate (Pn) and ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) activity increased, while activities of adenosine diphosphate glucose pyrophosphorylase (AGPase), sucrose synthase (SuSy) and sucrose phosphate synthase (SPS) decreased with more N under the same watering conditions. Compared to the plants without waterlogging, the leaf Pn and Rubisco activity, starch and sucrose contents of waterlogged rapeseed decreased at the two stages; activities of AGPase, SuSy and SPS of waterlogged rapeseed decreased at the seedling while increased at the bolting stage for both the two varieties. At the flowering stage, the Pn, the activities of Rubisco, AGPase, SuSy, SPS and contents of sucrose, starch increased with more N application for both ZS 9 and GH01. Compared to the plants without waterlogging, the Pn and Rubisco activity for waterlogged plants of the two varieties increased; the waterlogged plants of tolerant variety had higher activities of AGPase, SuSy and SPS, while those of sensitive variety was significantly lower. However, the decreased starch and sucrose content were found in both tolerant and sensitive varieties. The activities of AGPase, SuSy and SPS at flowering were highly positively correlated with yield under the interactive effects of N and waterlogging. These results suggested that the flowering stage is the most important stage that N had the positive regulation on waterlogged rapeseed growth. The carbohydrates translocation from leaves to seeds of the tolerant variety were enhanced after waterlogging, while that of the sensitive variety was still inhibited. This was the main reason for the difference in yield between the two waterlogged varieties.     

11个葡萄品种在乐山地区的花芽分化规律

研究了11个葡萄品种3年生树在四川乐山地区避雨栽培条件下的花芽分化表现。结果表明,结果母枝的花芽分化率,魏可、维多利亚、黑芭拉多、金手指、金优2号和巨玫瑰6个品种的都高于50%,较易形成花芽;户太8号、巨峰、夏黑的在43%~48%;美人指和红指较难形成花芽。结果母枝不同节位的花芽分化率,多数品种在第3节位≥50%;结果枝上花穗着生节位在第3.8~4.9节,其中双花率以巨玫瑰、巨峰和户太8号较高,分别为83.45%、73.61%和73.06%。以结果母枝的花芽分化率≥50%节位作为冬剪的最低节位,则金优2号、户太8号、巨玫瑰、金手指、维多利亚5个品种在第3~4节,黑芭拉多和巨峰2个品种在第2~3节,魏可在1~2节。夏黑表现花芽分化率较低,为43.33%,可能与夏黑不耐弱光有关,设施栽培时应加强透光性。     

粪肠球菌生长曲线的测定及其对小鼠脑组织的影响

为测定致羔羊脑炎粪肠球菌（Enterococcus faecalis）的生长曲线，寻求一种快速而准确的方法测定不同生长时期粪肠球菌数量，并客观评价其毒性强弱及其对小鼠脑组织的影响，试验采用平板菌落计数法和OD-Monitor振荡比浊法（D_λ值法）测定粪肠球菌的生长曲线，探究该菌在合适时间段内的吸光值（D_{600 nm}）与平板菌落计数法测定的活菌数（CFU）的关系。用粪肠球菌感染小鼠，观察记录小鼠的死亡情况，最后采用Karber法计算粪肠球菌感染小鼠的半数致死量（LD₅₀）。用LD₅₀的剂量感染小鼠，及时采集死亡小鼠脑组织，未死亡的小鼠72 h后全部剖杀取脑组织，一部分做涂片染色，制作病理切片，观察病理变化；一部分进行培养，用于PCR方法进行细菌的回收鉴定。结果显示，用两种方法测定此株粪肠球菌的生长曲线基本一致，在2~8 h生长迅速，为对数生长期，8~14 h生长缓慢，为稳定期，14 h之后死亡数增加，进入衰亡期；对12 h粪肠球菌D_{600 nm}与CFU的关系进行探讨，成功建立回归方程：y=20.769x-1.3422，R²=0.997；其感染小鼠的LD₅₀为7.77×10¹¹个活菌。以此剂量感染小鼠，脑组织涂片染色和培养染色，均能看到革兰氏阳性球菌；PCR结果显示，均出现了大小为112 bp的条带。对脑组织进行病理学观察发现该菌可导致脑组织充血、出血、形成微血栓，脑膜充血。通过生长曲线和其D_{600 nm}与CFU关系的建立，可实时监测粪肠球菌数量，为后期更深入研究粪肠球菌穿越血脑屏障的机制奠定重要的理论基础。     

正交设计优化青蒿SRAP-PCR反应体系及引物筛选

为了建立青蒿的SRAP最佳扩增体系,并筛选出SRAP多态性引物,本研究以青蒿叶片DNA为模板,采用正交试验设计,以Mg^2+、dNTP Mix、Taq DNA聚合酶、引物和DNA模板5种因素5个水平,对青蒿SRAP反应体系进行研究。结果表明,青蒿SRAP-PCR最佳反应体系为:引物0.6μmol/L、Mg^2+2.0 mmol/L、模板DNA 5.1 ng、Taq DNA聚合酶2.0 U、dNTPs 0.25 mmol/L,总体积为25μL。各因素对扩增反应均有不同影响,其中引物浓度的影响最大,dNTPs的影响最小。运用该体系对不同种质资源的青蒿进行验证,证明该体系稳定可靠,并在30个引物组合中筛选出了25对扩增条带清晰,多态性丰富的引物组合。这一结论为今后利用SRAP标记技术进行青蒿分子遗传学研究提供了科学依据。     

The microbial diversity and structure in peatland forest in Indonesia

The microbial community structure and function under forest in tropical peatlands are poorly understood. In this study, we investigated the microbial community structure and diversity in natural peat swamp forest soil, disturbed peat soil and mineral soil in Central Kalimantan, Indonesia, using 454 pyrosequencing. The results showed that the natural peat soil had the greatest fungal species richness (Chao1), which was significantly (p < .05) larger than that in the other two soils. Community structure of both fungi and bacteria in natural peat soil differed significantly from that in the disturbed peat soil (p = .039 and p = .045, respectively). Ascomycota (40.5%) was the most abundant phylum across the three soils followed by Basidiomycota (18.8%), Zygomycota (<0.1%) and Glomeromycota (<0.1%). The linear discriminant analysis with effect size (LEfSe) showed that Ascomycota (p < .05) and genus Gliocephalotrichum (p < .05) dominated in natural peat soil. Functionally, pathotrophs were more abundant in disturbed peat soil (p < .05). Proteobacteria (43.8%) were the most abundant phylum followed by Acidobacteria (32.6%), Actinobacteria (9.8%), Planctomycetes (1.7%). Methylocystis, Telmatospirillum, Syntrophobacter, Sorangium and Opitutus were the more abundant genera in disturbed peat soil, whereas Nevskia and Schlesneria were more abundant in mineral soil and natural peat soil, respectively. The natural peat forest soil supported a more diverse microbiology; however, the land use of such a soil can change its microbial community structure. The results provide evidence that the disturbance of tropical peat land could lead to the introduction and spread of a large number of fungal diseases     

Identification,characterization and full-length sequence analysis of a novel endornavirus in common sunflower (Helianthus annuus L.)

To identify the possible quarantine viruses in seven common sunflower varieties imported from the United States of America and the Netherlands, we tested total RNAs extracted from the leaf tissues using next-generation sequencing of small RNAs. After analysis of small RNA sequencing data, no any quarantine virus was found, but a double-stranded RNA(dsRNA) molecule showing typical genomic features of endornavirus was detected in two varieties, X3939 and SH1108. Full-length sequence and phylogenetic analysis showed that it is a novel endornavirus, temporarily named as Helianthus annuus alphaendornavirus(HaEV). Its full genome corresponds to a 14 662-bp dsRNA segment, including a 21-nt 5′ untranslated region(UTR), 3' UTR ending with the unique sequence CCCCCCCC and lacking a poly(A) tail. An open reading frame(ORF) that encodes a deduced 4 867 amino acids(aa) polyprotein with three domains: RdRP, Hel and UGT(UDP-glycosyltransferase). HaEV mainly distributed in the cytoplasm but less in the nucleus of leaf cells by fluorescence in situ hybridization(FISH) experiment. This virus has a high seed infection rate in the five varieties, X3907, X3939, A231, SH1108 and SR1320. To our knowledge, this is the first report about the virus of the family Endornaviridae in the common sunflower.     

优质国审棉品种——ZHM19

概述了ZHM19的选育过程、特征特性、产量、纤维品质和抗病性等特点及主要栽培技术措施。     

高产、抗病转基因抗虫棉品种——中棉所94A915

概述了中棉所94A915的选育经过、特征特性，并总结了其关键栽培技术。     

施用锌肥和遮阴互作对花生生长发育、抗病性及产量的影响

采用大田裂区试验，研究了施用锌肥和不同遮阴程度互作对花生生长发育、抗病性及产量的影响。结果表明，与不施锌肥相比，施用锌肥能提高花生不同部位锌含量、增加叶片SPAD值，提高花生叶片中可溶性糖、蛋白质和生长素含量，减少花生病害的发生，平均降低7.1个百分点，花生产量平均增加19.4%。相同施锌水平下，随着遮阴程度的增加，花生不同部位锌的含量和不同生育期叶片SPAD值以及花生叶片中可溶性糖、蛋白质和生长素含量呈增加趋势，花生的发病率比不遮阴对照增加4.8、10.2个百分点，花生产量平均降低16.5%、10.0%。在30%、70%的遮阴条件下，施用锌肥的花生产量比不施锌的分别提高21.1%、25.0%。本试验条件下，施用七水硫酸锌30 kg/hm2，使花生具有较强的抗低温寡照能力及抗病性能，增产显著，可在花生产区推广应用。