利用数码相机和Photoshop软件非破坏性测定叶面积的简便方法

采用数码相机获取叶片的数字图像,用Photoshop图像处理软件计算叶面积,并与目前常用的剪纸法和叶面积仪测定法进行比较分析。结果表明,本方法和上述传统测定方法测定结果存在极显著线性相关;不同拍摄分辨率、单位叶面积存贮像素个数和拍摄角度对测定结果无显著影响。和其它方法相比,本方法具有准确、快速、成本低廉、适合非破坏性动态连续观测等优点,适用于植物生理生态学研究中叶面积的测定。     

Identification and characterization of an NPV infection-related gene Bmsop2 in Bombyx mori L.

Abstract:  Using the fluorescent differential display technique, we analysed the differential expression of genes related to Bombyx mori nuclear polyhedrosis virus (BmNPV) resistance. Silkworm strains studied included the highly resistant strain NB, highly susceptible strain 306 and near-isogenic line 306NNZZ. One novel gene was identified and named Bmsop2 for its high similarity with the Sop2 protein of other species. It was identified to be linked to BmNPV susceptibility by Northern blotting and real-time polymerase chain reaction. The results indicated that it was actively expressed in midguts of strains 306, NB and the eighth generation of backcross (BC₈) of strain 306NNZZ which had been treated with BmNPV. But the expression level was low in the midguts of the control. In the mean time, the expression of Bmsop2 was the highest in strain 306 treated with BmNPV while it was the lowest in strain 306 not treated with BmNPV. Our study showed that Bmsop2 is a differentially expressed gene in strains NB, 306 and 306NNZZ which have different levels of resistance to BmNPV.     

Determination of baseline susceptibility to Cry1Ab protein for Asian corn borer (Lep., Crambidae)

Abstract: Although transgenic Bacillus thuringiensis (Bt) corn can provide a new tool for control of the Asian corn borer (ACB), Ostrinia furnacalis (Guenée), concern has been raised regarding the possibility of the target insect evolving resistance to the Bt protein under intensive selection pressure from Bt corn. Therefore, it is necessary to establish baseline data to enable detection of changes in susceptibility in field populations after prolonged exposure to Bt corn. Susceptibility to purified Cry1Ab protein from Bt was determined for 10 populations of ACB from the major corn‐growing regions of China, ranging geographically from Heilongjiang Province in the northeast to Shaanxi Province in the east‐central part. Neonate ACB were exposed to semi‐artificial diet incorporated with increasing Cry1Ab protein concentrations, and mortality and growth inhibition were evaluated after 7 days. The range of LC₅₀ (50% lethal concentration) among the populations was 0.10 to 0.81 μg/g (Cry1Ab protein/diet). Differences (P < 0.05) in susceptibility among the populations were significant. LC₅₀s generated from the Huanghuaihai Summer Corn Region were higher than those from the Spring Corn Regions. Bt was one of the significant natural biomortality factors of overwintering generation ACB. There was a significant correlation between percentage of the larvae infected with Bt and their LC₅₀ values to Cry1Ab protein in geographic distinct populations (r = 0.7350*, d.f. = 8, _r0.05 = 0.632). Based on the background of Bt formulations used for corn insect pests control in these areas, these differences were not caused by prior exposure to Bt insecticides. Instead, the small differences likely reflect natural Bt selection pressure. Because the variation in susceptibility to Cry1Ab was small (<10‐fold), the ACB apparently is susceptible to Cry1Ab across its range within China.     

A general method for nested RT-PCR amplification and sequencing the complete HCV genotype 1 open reading frame

Human sapovirus (SaV), an agent of human gastroenteritis, cannot be grown in cell culture, but expression of the recombinant capsid protein (rVP1) in a baculovirus expression system results in the formation of virus-like particles (VLPs). In this study we compared the time-course expression of two different SaV rVP1 constructs. One construct had the native sequence (Wt construct), whereas the other had two nucleotide point mutations in which one mutation caused an amino acid substitution and one was silent (MEG-1076 construct). While both constructs formed VLPs morphologically similar to native SaV, Northern blot analysis indicated that the MEG-1076 rVP1 mRNA had increased steady-state levels. Furthermore, Western blot analysis and an antigen enzyme-linked immunosorbent assay showed that the MEG-1076 construct had increased expression levels of rVP1 and yields of VLPs. Interestingly, the position of the mutated residue was strictly conserved residue among other human SaV strains, suggesting an important role for rVP1 expression.     

Kinetics of lipase-catalyzed hydrolysis of olive oil in AOT/isooctane reversed micelles

The kinetics of lipase-catalyzed hydrolysis of olive oil in AOT/isooctane reversed micellar media was studied. It was shown that the deactivation of lipase had a great influence on the reaction kinetics. Based on whether the enzyme deactivation and influences of both product and substrate on enzyme stability were included or not, four different kinetic models were established. The simulating results demonstrated that the kinetic model, which including product inhibition, enzyme deactivation and the improvements of lipase stability by both product and substrate, fit the experimental data best with an overall relative error of 4.68%.     

LGT蛋白质组阳性肿瘤患者免疫功能的观察报告——LGT系列论文之八

目的：了解LGT（lost goodwill target）蛋白质组阳性表达患者CD3^＋、CD4^＋、CD8^＋、CD4^＋/CD8^＋、T细胞和NK细胞的变化规律.方法：对30例LGT蛋白质组阳性表达的肿瘤患者分别采用美国BD公司生产的流式细胞检测仪及提供的相应单克隆抗体检测患者空腹血清CD3^＋、CD4^＋、CD8^＋、CD4^＋/CD8^＋、T细胞和NK细胞并用美国赛费吉（Ciphergen）公司制造的蛋白质指纹仪及该公司提供的弱阳离子交换芯片（WCX2）按其操作方法（SELDI检测技术）配对检测肿瘤患者空腹血清中的蛋白质指纹,对有病情加重的患者均检查两次以上.以指纹图上质荷比（M/Z）为11100＋H～11900＋H之间出现一峰簇样（cluster）的指纹标志为LGT阳性诊断标准,并按蛋白质指纹LGT检测阳性次数分成二组.对二组内CD3^＋、CD4^＋、CD8^＋、CD4^＋/CD8^＋、T细胞和NK细胞与正常参考值之间进行统计学显著性检验.结果：CD3^＋T细胞值在LGT蛋白质组持续阳性表达组是增高的,在另一组是无变化,二者之间有显著性差异,而CD8^＋T细胞在二组内同时增高,CD4^＋T细胞和NK细胞二组同时低下,无显著性差别.结论：本研究提示肿瘤晚期可能存在有酪氨酸蛋白激酶修饰的细胞内信号传导,使之病情加重,而肿瘤早期则不明显.这是一种新的看法,应加强这个方面的研究.     

Orchard floor management practices that maintain vegetative or biomass groundcover stimulate soil microbial activity and alter soil microbial community composition

Groundcover management systems (GMS) are important in managing fruit-tree orchards because of their effects on soil conditions, nutrient availability, tree growth and yields. We employed a polyphasic approach, incorporating measures of soil microbial abundance, activity and community composition, to study the long-term effects of different GMS on biotic and abiotic factors in an orchard soil. Four GMS treatments – Pre-emergence residual herbicides (Pre-H), post-emergence herbicide (Post-H), mowed-sod (Grass), and hardwood bark mulch (Mulch) – were established in 2-m-wide strips within tree rows in an apple orchard in 1992, and have been maintained and monitored annually until the present. We have measured soil water and nutrient availability, tree growth, and yields annually from 1993 to 2003. Soil nematode numbers and trophic groups were evaluated in July and Oct. 2001, and Sept. 2003. Numbers of culturable bacteria and fungi, soil respiratory activity, eubacterial and fungal community composition were determined in May and Sept. 2003. The Pre-H treatment soil had the fewest culturable bacteria, while the Grass treatment had the largest population of culturable fungi. Soil nematode population size and diversity were also affected by GMS treatments; the Pre-H treatment had the lowest ratio of (bacteriovores + fungivores) to plant parasitic nematodes. Soil respiration rates were higher in the Mulch than in other treatments during a 40-day incubation period. Hierarchical cluster dendrograms of denaturing gradient gel electrophoresis (DGGE) fingerprints for eubacterial community 16S rRNA genes indicated that Post-H and Grass treatments clustered together and separately from the Pre-H and Mulch treatments, which were also grouped together. The influence of GMSs on the fungal community, as assessed by PCR-DGGE of the internal transcribed spacer (ITS) region, was not as pronounced as that observed for bacteria. Soil fungal community composition under the Mulch differed from that under other treatments. The effects of GMS on soil microbial community abundance, activity, and composition were associated with observed differences in soil organic matter inputs and turnover, nutrient availability, and apple tree growth and yields under the different GMS treatments.     

RUS家族对植物生长发育的调控作用

叶绿体基因组编码许多参与光合作用和其他代谢过程的关键蛋白质,在叶绿体中合成的代谢物对于植物正常的生长发育至关重要。根对紫外线-B辐射敏感[Root-UVB (ultraviolet radiation B)-sensitive, RUS]蛋白属于叶绿体蛋白,由高度保守的DUF647结构域组成,在参与植物形态发生、物质运输和能量代谢等多种生命活动的调控中发挥作用。本文就近年来关于RUS家族在植物的胚胎发育、光形态建成、维生素B6稳态、生长素转运和花药发育等生长发育过程中的相关研究进行回顾和总结,为深入研究其在植物生长发育中的分子调控机制提供了参考。     

一株抗茶炭疽菌和魔芋镰刀菌的淀粉酶产色链霉菌的分离、鉴定及生防潜能

【背景】放线菌是一类重要的生防菌,具有强大的代谢活性,能产生抗生素、酶、酶抑制剂和激素等天然产物抑制病原物生长。【目的】从茶树根际分离得到放线菌,研究候选放线菌对茶炭疽菌和魔芋镰刀菌的抑菌活性及其生防潜能。【方法】分别以茶炭疽病致病菌Colletotrichum camelliae和魔芋茎腐病致病菌Fusarium solani为指示菌,采用土壤稀释涂布法、平板对峙法和菌丝生长速率法,从茶树根际土壤中分离、筛选拮抗放线菌,并根据菌株的形态特征、生理生化特性和系统发育分析结果对其进行分类鉴定,并开展候选放线菌的产促生相关物质和分泌细胞壁水解酶能力的定性检测试验。【结果】共分离得到14株拮抗放线菌,菌株A-dyzsc04-2的拮抗效果最强,被鉴定为淀粉酶产色链霉菌（Streptomyces diastatochromogenes）。该菌株的活菌体对C. camelliae和F. solani的抑制率分别为66.71%±1.23%和71.59%±2.46%,其无菌发酵滤液对2种指示菌的抑菌率均大于90%;此外,菌株A-dyzsc04-2还具有产嗜铁素和葡聚糖酶以及溶解无机磷的能力。【结论】菌株A-dyzsc04-2是一株优良的生防菌,具有较高的开发利用价值,研究结果为菌株A-dyzsc04-2防治茶炭疽病和魔芋茎腐病提供了理论支撑。