不同催青方式对二化性家蚕过氧化氢酶基因表达的影响

25℃明催青和15℃暗催青分别诱导二化性家蚕Bombyx mori 产滞育性卵和非滞育性卵。此前我们的研究表明, 上述催青处理的二化性家蚕H₂O₂水平存在显著差异。过氧化氢酶（catalase, CAT）是昆虫清除H₂O₂的关键酶。为了进一步明确家蚕滞育过程中H₂O₂代谢的调控机制, 用RT-PCR测定了上述两种催青处理对二化性家蚕CAT基因表达的影响。结果表明：25℃明催青显著提高了滞育诱导和决定阶段的CAT mRNA 水平和CAT活性。滞育性卵的CAT mRNA水平在产后24 h形成峰值, 在72 h后消失; CAT活性在96 h前上升, 120 h后保持于低水平。非滞育性卵的CAT mRNA水平和CAT活性都随着胚胎发育而上升。可见, 25℃明催青诱导二化性家蚕子代滞育可能是通过影响CAT基因表达来调节H₂O₂水平。     

家蚕二化性杂交品种部分数量性状遗传参数的初步评估

采用完全随机设计法根据10头老熟幼虫体重、全茧重、茧层量、茧层率（%）、存活率、万蚕茧层量和茧丝长等指标,对两对二化性家蚕Bombyx mori L. 杂交品系（SH6×NB₄D₂和CSR₂×CSR₄）杂交一代的22个子代个体进行了遗传参数估算,以缩小优质蚕品种的候选范围,并且计算出直接筛选的参数,如遗传力和遗传进度等,使这些信息可用于以筛选高产新品种为目的的育种和选择过程中。杂交子代2, 4, 5, 6, 7, 10, 14, 16, 19和20号个体在这几个指标中表现出显著的优越性。全茧重、万蚕茧层量和茧丝长的遗传力和遗传进度较大,可以简单地从表现型的差异对这些性状进行选择并取得遗传性状改良。其他几个指标(10头老熟幼虫体重、茧层量、茧层率(%)和存活率)的遗传力和遗传进度较低,对这些性状进行直接选择来改良品种的效果较差。     

不同水分含量桑叶对二化性家蚕的营养效应

研究了取食不同水分含量桑叶对二化性家蚕(CSR3×CSR6)5龄幼虫的影响。实验设嫩叶(水分含量80%～85%)、半成熟叶(水分含量65%～70%)和成熟叶(水分含量55%～60%)3个不同处理。采用标准重量分析法分析和计算各种生长、营养和营养效率指数。结果表明,在取食较高水分含量桑叶的处理中,营养指数(包括取食量、消化量和近似消化率等)和营养效率指数(包括食物利用率和转化率等)均显著较高,但不同处理中消化食物向茧壳的转化率及产生每克茧壳所需食物消化量差异不明显。结论是取食水分含量高的桑树嫩叶有助于提高家蚕的取食和营养。     

二化性家蚕滞育、非滞育和即时浸酸卵中糖代谢相关酶mRNA水平和活性的变化

【目的】探究家蚕Bombyx mori胚胎发育早期卵内糖代谢与滞育的关系。【方法】以家蚕二化性品种"秋丰"活化越年卵为材料,一部分于17℃暗催青,孵化后常规饲养,制备非滞育命运卵(ND);另一部分于25℃明催青,孵化后常规饲养,制备滞育命运卵(DD),再用盐酸溶液处理部分DD卵制备即时浸酸卵(IA)。在卵产下或浸酸后0, 2, 6, 12, 24, 48, 72和96 h分别取样,利用qRT-PCR测定家蚕卵中己糖激酶(hexokinase, HK)、磷酸果糖激酶(phosphofructokinase, PFK)、山梨醇脱氢酶(sorbitol dehydrogenase, SDH)、磷酸烯醇式丙酮酸羧激酶(phosphoenolpyruvate carboxykinase, PEPCK)和海藻糖酶(trehalase, TRE)5种糖代谢相关酶基因的mRNA表达水平,利用紫外-可见分光光度法测定家蚕卵中这5种酶的活性。【结果】家蚕糖酵解途径关键酶己糖激酶(BmHK)和磷酸果糖激酶(BmPFK),以及糖分解代谢关键酶山梨醇脱氢酶(BmSDH-1)和海藻糖酶(BmTRE)在ND和IA中...     

氟化物对家蚕耐氟和氟敏感品种肠道中留存产酶菌群落组成和多样性的影响

【目的】研究氟化物对家蚕Bombyx mori肠道内留存产酶菌的影响, 有助于了解家蚕耐氟和氟敏品种的耐氟力差异。【方法】分别给家蚕耐氟品种T6和氟敏品种734添食NaF溶液浸泡后的新鲜桑叶, 至5龄第3天取材。采用筛选培养基筛选产蛋白酶、 纤维素酶、 脂肪酶、 淀粉酶的菌株, 并结合16S rDNA系统发育关系, 对菌株进行分类鉴定。【结果】家蚕肠道内产消化酶细菌有芽孢杆菌属Bacillus、 葡萄球菌属Staphylococcus、 肠杆菌属Enterobacter和不动杆菌属Acinetobacter和微小杆菌属Exiguobaterium, 其中芽孢杆菌Bacillus sp.、 葡萄球菌Staphylococcus sp. 和不动杆菌Acinetobacter sp.细菌可同时产4种酶。氟中毒后T6肠道内产酶菌由5种减少到4种, 734肠道内产酶菌由2种减少到1种。【结论】家蚕肠道内留存的产酶菌与家蚕自身的耐氟能力相关。     

利用二代测序技术高通量分析水稻品种间抗病性差异的分子基础

稻瘟病和白叶枯病是由稻温病菌(Magnaporthe oryzae)和白叶枯病菌(Xanthomonas oryzae pv.oryzae)引起的两种主要水稻病害,也是制约中国水稻生产的主要病害。为了从DNA水平探索造成水稻感病品种‘丽江新团黑谷’(LTH)和高抗品种‘特特普’(Tetep,TTP)间抗病性差异的分子基础,该研究对其已知的3个抗稻瘟病基因和3个抗白叶枯基因所在DNA区段分别进行PCR扩增,将等量混合的PCR产物再与基因组重测序样品按Ct值差值(ΔCt)~10的比例混合,采用二代测序技术进行一次性测序和比较分析,并对有差异的基因区域进行常规传统测序验证,以确定这2个品种中抗性基因(R基因)的数目和结构与品种抗病或感病表型的关联性。实验结果表明,二代测序能够快速并准确地寻找到2个不同水稻品种中多个特定基因的序列差异,且差异位点与常规测序结果相符。从LTH和TTP这2种抗性不同水稻品种在多个抗性基因的DNA水平差异来看,有差异的抗性基因位点在高抗品种TTP中大都与原始抗性基因序列相同,而对应的普感品种LTH的抗性基因往往多表现为氨基酸突变,这些序列差异很可能就是导致TTP与LTH抗性差异的分子基础。     

鲫鲤杂种一代(F_1)自交二代(F_2)的受精细胞学研究

荷包红鲫 (♀ )×湘江野鲤 (♂ )杂交产生的杂种一代 ( F1)成熟卵的直径为1 .0～ 1 .2 5mm;为单受精孔卵 ,一般只允许单个精子入卵 ;成熟卵在受精前处于第二次减数分裂中期 ;鲫鲤杂种一代的雄性部分可育 ,能产生正常的精子 ;杂种一代的受精方式为单精受精 ,具有正常的受精细胞学程序 .受精 30 s后 ,精子通过受精孔进入卵质 ;5min后 ,精子产生明显星光 ;1 5min后 ,精子头部膨大核化 ,最后形成雄性原核 ,与此同时 ,雌性原核也开始形成 ;2 5min后 ,雌、雄原核向胚盘中央靠近 ,然后彼此接触 ,最后融合为合子核 ;40 min后 ,开始第一次卵裂 ,在一个受精卵第一次卵裂中期的切片上看到了一个三极纺锤体 .     

二斑叶螨对不同甜瓜品种的取食和产卵选择性

二斑叶螨Tetranychus urticae Koch是世界性的重大农业害螨,为明确其对不同甜瓜品种的取食和产卵选择性,采用离体叶碟法研究了二斑叶螨在7种不同甜瓜品种上的选择性及其产卵量,并探究了其在活体植株上的种群增长动态。结果表明,离体叶碟上,二斑叶螨对不同甜瓜品种的取食和产卵选择存在明显差异。选择2 h时二斑叶螨在IVF29品种上的成螨数量最低,而在IVF168上成螨数量最高,选择率为28.73%;选择12 h时,二斑叶螨成螨量及产卵量在IVF58和IVF118品种上较低,而在羊角蜜和IVF168品种上表现较高,24 h时选择趋势一致。活体植株上的种群动态结果表明,调查时间段内二斑叶螨种群数量持续升高,第20天时羊角蜜、IVF58和IVF118种群数量(低于400头/株)显著低于IVF117和IVF168的螨量(高于660头/株)。对测试不同时间段内各个参数进行综合比较和分析,二斑叶螨对甜瓜IVF168品种的选择性强、适合度高,而对IVF58和IVF118品种的选择性较差,表明IVF58和IVF118可用于甜瓜生产中的备选品种及高抗螨性甜瓜新品种的选育。     

二斑叶螨对不同品种茄子的选择性与适合度

【目的】为明确二斑叶螨Tetranychus urticae Koch对不同品种茄子的选择性与适合度。【方法】通过二斑叶螨对不同品种茄子的田间选择性试验以及在室内不同品种茄子对二斑叶螨种群数量、个体生长发育和繁殖的影响试验,分析了二斑叶螨对9个茄子栽培品种和1个野生茄子品种的寄主选择性,以及在不同品种上的种群动态、发育历期和繁殖力。【结果】二斑叶螨对野生品种蒜芥茄Solanum sisymbriifolium的选择性最差,在该品种上除卵可以正常孵化外其他时期均不能存活。在茄子S.melongena品种间,二斑叶螨对京茄6号和日友长直壮青长茄的选择性最强,分别达到195和183头/株,对布利塔和长竹丝茄的选择性最差,分别为63.5和59.75头/株。接入20对成螨后第28天,紫龙王4号上的数量最多(1 009头/株),京茄6号和嫁接的京茄6号上的数量次之(分别为981和909头/株)。二斑叶螨在京茄6号上的发育历期最短,为9.31 d;在长丰2号上发育历期最长,为11.08 d。京茄6号、京茄黑霸和嫁接的京茄6号上二斑叶螨开始产卵的前5 d内单雌产卵量分别为35.92,33.20和31.34粒/头,是产卵量较多的3个品种,且3个品种之间差异不显著;紫龙王4号、茄砧1号、布利塔和长丰2号上的产卵量较少,分别为18.56,19.24,22.26和23.36粒/头,且4个品种之间差异不显著。【结论】二斑叶螨对紫黑圆茄类的京茄6号的选择性较强、适合度较高,对紫黑长茄布利塔的选择性和适合度为中等,对野生品种蒜芥茄的选择性和适合度表现为最差。     

一种基于二代测序辨识短串联重复序列长度变异的新方法及其在人类遗传疾病研究中的应用

二代测序技术的涌现推动了基因组学研究,特别是在疾病相关的遗传变异研究中发挥了重要作用．虽然大多数遗传变异类型都可以借助于各种二代测序分析工具进行检测,但是仍然存在局限性,比如短串联重复序列的长度变异．许多遗传疾病是由短串联重复序列的长度扩张导致的,尤其是亨廷顿病等多种神经系统疾病．然而,现在几乎没有工具能够利用二代测序检测长度大于测序读长的短串联重复序列变异．为了突破这一限制,我们开发了一个全新的方法,该方法基于双末端二代测序辨识短串联重复序列长度变异,并可估计其扩张长度,将其应用于一项基于全外显子组测序的运动神经元疾病临床研究中,成功地鉴定出致病的短串联重复序列长度扩张．该方法首次原创性地利用测序读长覆盖深度特征来解决短串联重复序列变异检测问题,在人类遗传疾病研究中具有广泛的应用价值,并且对于其他二代测序分析方法的开发具有启发性意义．     

羊草与灰色赖草F1代细胞悬浮系的建立及植株再生

本研究以羊草(L eym us ch inensis)与灰色赖草(L eym us cinereus)杂种F1代幼穗为外植体诱导愈伤组织,在3.0 m g/L 2,4-D M S培养基上继代1次后,转入不同浓度激素(2,4-D、IAA、KT)配比和不同浓度蔗糖的M S液体培养基进行振荡培养,建立杂种F1代细胞悬浮系和植株再生体系.结果表明,细胞悬浮培养时,M S 1.0 m g/L2,4-D 0.1 m g/L KT 4%蔗糖的液体培养基最佳;悬浮细胞分化时,1.0 m g/L 2,4-D 0.1 m g/L KT 4%蔗糖 M S和1.0 m g/L 2,4-D 4%蔗糖 M S培养的悬浮细胞在1.0 m g/L NAA 0.5 m g/L KT M S分化培养基上的绿苗分化率分别达到83%和80%.细胞悬浮系及再生体系的建立为杂种F1代育性恢复的研究奠定了基础.     

Rearing of silkworm larva in a closed gas revitalizing plant system

Plant growing and insect breeding experiments in the CICS (Closed Integrative Cultivating System) were carried out. The CICS was established for collecting experimental data to investigate gas circulation and mass exchange between plants and animals as well as animal growth and nutrient compositions in the Bioregenerative Life Support System. The CICS was 1.4 m high with the base measuring 1.4 m × 0.8 m. In the plant chamber, stem lettuce plants were grown in a staggered manner. Silkworms in five different instars were fed in the animal chamber. Every 4 days, the silkworms that had been in the 5th instar for 3 days and all the silkworm’ excreta were taken out of the system. Meanwhile, the silkworms in the first instar were transferred into the animal chamber. During this process, O₂ and CO₂ concentrations in the CICS were measured, and the growth and nutrient composition of the silkworms in the system were compared with those of silkworms reared in the open environment. Results showed that O₂ and CO₂ concentrations in the system were 19.07-20.61% and 0.11-0.35%, respectively. The comparison of growing differences between the silkworms fed in the animal chamber and those in the open environment indicates the insect's biomass increasing rates and bioconversion rates in the CICS were lower than those in the open environment, and the amounts of excreta produced in the animal chamber were larger. Protein content of the SP (Silkworm Powder) produced in the CICS was more than that in the open environment, while fat content of the SP in the CICS was lower than that in the open environment. Calcium, phosphorus and iron contents of the SP produced in the CICS were 139.00 mg calcium/100 g SP, 1.20 mg phosphorous/100 g SP, and 7.95 mg iron/100 g SP. In terms of amino acids, the quality of the SP produced in the CICS was equivalent to that gained in the open environment.     

Predicting the potential of open-pollinating populations for the production of superior F1 hybrids

Summary The distributive properties of a single population or of a population resulting from a cross between two populations are reproduced when inbreds randomly extracted from the population itself or from the two parental populations are randomly paired. Hence, population parameters that are usually obtained during a breeding programme can be used to predict the performance of the F₁ hybrids that can be derived from them at that stage. Multiple allelism, epistasis and deviations from Hardy-Weinberg equilibria should not cause biasis to the predictions. While in theory genotype x environment interaction and linkage disequilibrium may disturb the predictions, in practice they are unlikely to create problems that cannot be accommodated. Genotypic and phenotypic predictions of the proportions of the F₁ hybrid distribution scoring above or below a given standard are made and analysed for three characters, weight of the ears, plant height and height of the ear, in two populations of maize per se and their interpopulational cross. Because no random inbred lines from the experimental populations are presently available we cannot check our predictions. However, genotypic and phenotypic predictions and observations of F₁ hybrids obtained from populations created by computer simulation are provided to illustrate our procedures.J. F. F. de Toledo is a geneticist on leave from Centro National de Pesquisa de Soja-EMBRAPA, Caixa Postal 1061, 86100-Londrina-Pr, BrazilJ. B. de Miranda Filho is Professor of Genetics at Escola Superior de Agriculture Luiz de Queiroz-ESALQ, Caixa Postal 83, 13400-Piracicaba-SP, Brazil     

Phylogenetic analysis of H1N1 sequences from pandemic infections during 2009 in India

Since April 2009, a serious pandemic infection has been rapidly spread across the world. These infections are caused due to the novel swine origin influenza A (H1N1) virus and hence these are commonly called as "Swine Flu". This new virus is the reassortment of avian, human and swine influenza viruses and thus it has a unique genome composition. There are 16 different types of hemagglutinin (HA) and 9 different types of neuraminidase (NA) that can be genetically and antigenetically differentiated. The first influenza A virus isolated from pigs was of the H1N1 subtype and these viruses have been reported to cause infection in pigs in many countries. The outbreak of this virus has been transmitted from pigs to humans. This new reassorted (exchange of genes) virus which is the cause of 2009 pandemic infections has the ability to spread from human to human. This spread of infection should be brought to an end. In this study, a phylogenetic analysis of the nucleotide sequences of the RNA segments of human H1N1 viruses was carried using MEGA version 4.0 to demonstrate the route map of infection to India. Phylogenetic analysis of the sequences from India, published in Influenza Virus Resource (a database that integrates information gathered from the Influenza Genome Sequencing Project of the National Institute of Allergy and Infectious diseases (NIAID) and the genbank of the (NCBI)) was retrieved and used for the analysis. The results showed that the various segments of the Indian isolates clustered well with the sequences from American, Asian and European countries and thus indicating the transmission of viruses from these places to India.     

利用杉木的F1代群体构建遗传连锁图谱

对于杉木 1∶1分离的分子标记位点 ,提出了一种新的构建遗传连锁图谱的策略。通过二点连锁分析 ,任意两个位点的连锁相和重组率可以得到推断和估计。对于一个连锁群中的最优排序 ,采用隐马尔可夫链模型的方法进行多位点的连锁分析。该作图方法比通常林木上所用的“拟测交”作图方法更有效。采用该作图策略 ,利用句容0号无性系 (♀ )×柔叶杉 (♂ )的F1代群体的AFLP分子标记数据重建了句容 0号无性系和柔叶杉的遗传连锁图谱。在句容 0号无性系的连锁图谱中 ,有 10 1个标记分布在 11个连锁群上 ,图谱的总长度为 2 2 82 6cM ,平均图距为 2 2 6cM ,单个连锁群上最多含有 17个标记 ,最少含有 5个标记 ;在柔叶杉的连锁图谱中 ,有 94个标记分布在 11个连锁群上 ,图谱的总长度为 2 5 6 5 8cM ,平均图距为 2 7 3cM ,单个连锁群上最多含有 16个标记 ,最少含有 4个标记。构建的句容 0号无性系和柔叶杉的遗传连锁图谱比原有的图谱分别增加了 2 6个标记和 2 8个标记 ,双亲的图谱共增加了 5 4个AFLP标记 ,使图谱上的分子标记总数达到 195个 ,双亲遗传图谱的跨度均超过了 2 0 0 0cM ,基本上达到了杉木基因组的长度 ,图谱的覆盖率接近于 10 0 %。利用新的作图方法可以较大提高分子标记在图谱上的分辨率 ,得到可认为是     

利用杉木的F1代群体构建遗传连锁图谱

对于杉木11分离的分子标记位点,提出了一种新的构建遗传连锁图谱的策略.通过二点连锁分析,任意两个位点的连锁相和重组率可以得到推断和估计.对于一个连锁群中的最优排序,采用隐马尔可夫链模型的方法进行多位点的连锁分析.该作图方法比通常林木上所用的"拟测交"作图方法更有效.采用该作图策略,利用句容0号无性系(♀)×柔叶杉(♂)的F1代群体的AFLP分子标记数据重建了句容0号无性系和柔叶杉的遗传连锁图谱.在句容0号无性系的连锁图谱中,有101个标记分布在11个连锁群上,图谱的总长度为2 282.6 cM,平均图距为22.6 cM,单个连锁群上最多含有17个标记,最少含有5个标记;在柔叶杉的连锁图谱中,有94个标记分布在11个连锁群上,图谱的总长度为2 565.8 cM,平均图距为27.3 cM,单个连锁群上最多含有16个标记,最少含有4个标记.构建的句容0号无性系和柔叶杉的遗传连锁图谱比原有的图谱分别增加了26个标记和28个标记,双亲的图谱共增加了54个AFLP标记,使图谱上的分子标记总数达到195个,双亲遗传图谱的跨度均超过了2 000 cM,基本上达到了杉木基因组的长度,图谱的覆盖率接近于100%.利用新的作图方法可以较大提高分子标记在图谱上的分辨率,得到可认为是覆盖了整个基因组的遗传连锁框架图.     

Resistance of tomato F1 hybrids to grey mold

A two-year estimation of Botrytis cinerea stem damage of 299 experimental F₁hybrids grown in 2 rotations: winter-autumn and summer-autumn was carried out. The estimation was made in a period of mass fruitage. As a result it was shown, that the hybrids with determinate type of growth (sp) are damaged more strongly than indeterminate ones (sp +). An average disease severity in the first group was 2.8 against 2.3 in the second one using a scale 0–4. Number of strongly injured (2.5–4) plants in each group were 44.1 % and 30.7 % and number of moderate resistant (0–1.5 numbers) plants were 10 % and 8.5 % accordingly. The analysis of resistance of F₁ hybrids determined significant influence of parental lines on a degree of a damage of plant stems by B. cinerea.     

Multivariate analysis of tooth crown morphology in Japanese-American F1 hybrids

Mesiodistal crown diameters of I1 through M1 and six non-metric crown traits in permanent dentition of Japanese-American F₁ hybrids were compared with those of the parental populations. The hybrids were born of Japanese females and American males, both Caucasians and American Blacks, after World War II and brought up at Elizabeth Saunders Home in Kanagawa Prefecture, Japan. The comparisons were undertaken by means of multivariate analysis methods such as principal component analysis, distance and similarity coefficients and multidimensional scaling. The F₁ hybrids generally occupy an intermediate position of the two parental populations, and this is particularly evident in size component of the crown measurements and in distance analysis of frequencies of the non-metric crown traits. The shape component of the crown measurements, however, not necessarily follows such a rule and suggests a more complicated gene control than in the case of the size component. Also, each measurement and frequency of non-metric trait of tooth crowns in F₁ hybrids is not uniformly at the middle position between their parental populations but this is true when they are analyzed as multivariables.