Bulk pollen pollination in maize for efficient construction of introgression populations with high genome coverage

Introgression populations consist of a set of introgression lines or families, constructed by continuous backcrossing to the recurrent parent, while carrying a limited number of chromosome segments from a donor parent in their genomes. Increasing the genome coverage is an important aim when constructing introgression population. In this study, we proposed bulk pollen pollination (BPP) method and used it to increase the genome coverage of a maize introgression population. The results showed that the genome coverage of the introgression population constructed using BPP method reached 100% at BC₃ generation, which accorded with the simulation result. The BPP‐based BC₃F_1:2 population could identify most quantitative trait loci (QTL) detected using the F_2:3 population, especially major QTL. Simulation analysis showed that the genome coverage of introgression population increased with the increase of population size and the number of bulked plants, and decreased with the increase of backcross generation. Our results proved the reliability of the BPP‐based introgression population in increasing genome coverage and detecting QTL, and provided references for constructing high‐coverage introgression populations.     

香蕉与B基因组相关的SCAR标记研究

香蕉(Musa L.)是由2个二倍体野生种Musa acuminata Colla(AA基因型)和Musa balbisiana Colla(BB基因型)种内或种间杂交进化而来,其B基因组中带有重要的优良基因。利用与香蕉B基因组相关的gypsy-IRAP分子标记,成功开发了一对SCAR引物,适用于鉴定尖叶蕉(AAw)、长梗蕉(BB)、香牙蕉(AAA)、贡蕉(AAcv)、大蕉、粉蕉(ABB)、粉大蕉(ABB)、龙牙蕉(AAB)以及四倍体香蕉(AAAB)等是否含有B基因组。     

Pigeonpea improvement: An amalgam of breeding and genomic research

In the past five decades, constant research has been directed towards yield improvement in pigeonpea resulting in the deployment of several commercially acceptable cultivars in India. Though, the genesis of hybrid technology, the biggest breakthrough, enigma of stagnant productivity still remains unsolved. To sort this productivity disparity, genomic research along with conventional breeding was successfully initiated at ICRISAT. It endowed ample genomic resource providing insight in the pigeonpea genome combating production constraints in a precise and speedy manner. The availability of the draft genome sequence with a large‐scale marker resource, oriented the research towards trait mapping for flowering time, determinacy, fertility restoration, yield attributing traits and photo‐insensitivity. Defined core and mini‐core collection, still eased the pigeonpea breeding being accessible for existing genetic diversity and developing stress resistance. Modern genomic tools like next‐generation sequencing, genome‐wide selection helping in the appraisal of selection efficiency is leading towards next‐generation breeding, an awaited milestone in pigeonpea genetic enhancement. This paper emphasizes the ongoing genetic improvement in pigeonpea with an amalgam of conventional breeding as well as genomic research.     

大花君子兰叶绿体基因组及其特征

采用Illumina MiSeq测序平台对大花君子兰（Clivia miniata）叶片总DNA进行测序，通过组装获得了其叶绿体基因组（cpDNA）全长序列（158 114 bp）。对其cpDNA注释得到135个基因，包含87个蛋白编码基因、40个tRNA基因和8个rRNA基因。采用生物信息学方法对获得的cpDNA进行简单序列重复（SSR）分析和密码子偏好性分析。结果显示：①大花君子兰cpDNA中共有61个SSR位点，其中单核苷酸、二核苷酸、三核苷酸、四核苷酸、五核苷酸和六核苷酸重复数分别为38、9、2、8、3和1个，多数SSR分布在基因间隔区；②大花君子兰cpDNA密码子偏爱以A或U（T）结尾，亮氨酸使用频率最高，半胱氨酸使用频率最低。基于24种植物的cpDNA全长和23种植物的叶绿体ycf2基因序列进行系统发育分析，结果显示大花君子兰与石蒜科植物在同一分支，显示最近的亲缘关系，支持大花君子兰属于石蒜科。基于叶绿体ycf2的系统发育分析结果与基于cpDNA全长的系统发育分析研究结果大部分相同，支持ycf2基因可以代替cpDNA全长用于植物系统发育分析。     

热风干燥过程中山药水分状态的变化研究

为了解山药的水分赋存状态的变化，以襄阳道地山药为材料，分别在60、70、80、90、100 ℃条件下进行热风干燥，采用低场核磁共振（LF-NMR）和差式量热扫描（DSC）技术，每隔15 min测定山药在热风干燥过程中的水分状态及迁移规律。结果表明，在60~90 ℃条件下，温度越高，干基含水率的降速越快。山药的T2弛豫图谱有3个较为明显的吸收峰，随干燥进程的延续，各峰面积均明显减少，其中自由水所在峰的面积降幅最大，表明干燥过程中自由水散失最多，而且自由水逐渐向半结合水和结合水迁移，冷冻峰和解冻峰也随之变小。但在100 ℃下干燥时，样品可能因表面板结导致干基含水率、低场核磁吸收峰升高，冷冻和解冻峰面积增加。因此，在实际干燥过程中，山药的热风干燥温度不宜高至100 ℃。     

THE EFFECT OF A COMBINATION OF MEDETOMIDINE-BUTORPHANOL AND MEDETOMIDINE, BUTORPHANOL, ATROPINE ON GLOMERULAR FILTRATION RATE IN DOGS

The effects of intramuscularly administered medetomidine and butorphanol (MB), and medetomidine, butorphanol, atropine (MBA) on glomerular filtration rate (GFR) were determined in six dogs as measured by 99m-Tc-labeled diethylenetriaminepentaacetic acid (99mTc-DTPA) nuclear scintigraphy. Direct systolic, diastolic, and mean arterial blood pressures and heart rate were measured at regular time intervals before, during, and after GFR calculations. The mean GFR measurement following MB was significantly greater (4.44 ml/min/kg) than following MBA (3.82 ml/min/kg) or saline treatment (3.41 ml/min/kg). There was no significant difference between the mean GFR measurements following MBA injection and following saline injection. Diastolic and mean arterial pressures following MBA injection were significantly higher than the values recorded after either MB or saline alone. Heart rate following MB administration was significantly lower than that recorded for dogs receiving MBA or saline alone. The results of this study indicate that the administration of medetomidine in combination with butorphanol significantly increases total GFR in healthy dogs, while the administration of the combination of medetomidine, butorphanol, and atropine does not.     

转基因动物研究进展

转基因动物是现代生物技术中一个极其重要的研究领域，目前已经有转基因小鼠、兔、绵羊、山羊、猪、牛、鸡和鱼等多种转基因动物问世。本文综述了转基因动物的制作方法、转基因动物的应用研究以及所取得的重要成就，并指出了转基因动物存在的主要问题，展望了其发展前景。     

影响体细胞核移植效率的因素

哺乳动物体细胞核移植 (克隆 )是近年来迅速发展起来的一项新技术 ,对濒危动物保护、优良种畜快速扩群以及核质关系的研究、药物试验等诸多领域都具有重大意义。但体细胞核移植是一项程序繁杂的工作 ,每一步操作所采用的试剂、材料和方案等都会影响到核移植效率 ,因而其影响因素十分众多。文章根据目前已有资料 ,论述了卵母细胞来源、卵母细胞与供体细胞的胞质容量、卵母细胞的去核方案、体细胞供体动物的年龄、供体细胞的组织来源与传代次数及供体细胞的冷藏、冷冻和所处的细胞周期阶段对核移植效率的影响。另外 ,对重组胚的融合 -激活方案、核移植方案及重构胚的培养方案对核移植效率的影响也作了简要论述。     

BmNPV半胱氨酸蛋白酶基因的核苷酸序列研究

对家蚕核型多角体病毒苏州株（BmNPVsu）光胱氨酸蛋白酶基因（CP）的序列分析表明,该基因读码框为972个核苷酸,编码323个氨基酸。同源性分析表明,BmNPVsu的CP与首蓿银纹夜蛾核型多角体病毒（AcNPV）、美国白蛾核型多角体病毒（HcNPV）、云杉卷叶蛾核型多角体病毒（CfNPV）、黄杉毒蛾核型多角体病毒（OpNPV）、舞毒蛾核型多角体病毒（Ld-NPV）在DNA水平上的同源性分别为96.5％、76.2％、74.9％、72.7％、62.9％;在氨基酸水平上的同源性分别为96.9％、77.1％、79.3％、77.1％、65.6％。BmNPV CP的氨基酸序列与不同来源的木瓜蛋白酶超家族的CP也具有较高的同源性,特别与 Trpanosoma brucei的CP具有较高的一致性,达32％。在组织蛋白酶B、H、L、S以及木瓜蛋白酶的36个保守氨基酸残基中有31种出现在BmNPVsu的CP中,BmNPVsu CP同其它杆状病毒CP一样,可看作木瓜蛋白酶超家族成员。     

Nuclear transfer in loach (Paramisgurnus dabryanus Sauvage) by cell-to-cell electrofusion

To study nuclear transfer in the loach (Paramisgurnus dabryanus Sauvage), blastula and gastrula cells were fused with UV-inactivated oocytes by cell-to-cell electrofusion. To facilitate nuclear transfer, blastula and gastrula cells were cultured or incubated at 4 °C in different solutions. TC-199 medium supplemented with 20% calf serum was the best culture solution, and effectively retained the totipotence of blastula or gastrula cells for up to 10 days. It was found that gastrula cells incubated at 4 °C had the same totipotence as blastula cells. The optimal UV dosage for inactivation of the oocyte chromatin was 180–240 mJ cm⁻². Electrofusion was carried out in a cone-shaped fusion chamber, which permitted the recipient oocyte and the donor blastula cell to contact one another. The electrofusion procedure resulted in a 10% success rate of normal-appearing fish. Genetic analysis indicated that the nuclear material originated from the donor cell (blastomere) and the oocyte pronucleus did not take part in development.