油菜黑胫病菌T-DNA插入诱变因素优化及突变体筛选

油菜黑胫病是由Leptosphaeria biglobosa引起的一种真菌病害。这种病害在我国油菜产区广泛发生，并造成一定的经济损失。为通过获取突变体来研究L. biglobosa的生态适应性机制及致病机制，本文优化了影响农杆菌介导转化（ATMT）油菜黑胫病菌L. biglobosa菌株Lb731的因素，评估转化子质量，并筛选相关突变体。结果明确了农杆菌介导转化菌株Lb731的最佳因素：潮霉素B浓度为50 μg/mL，转化受体（分生孢子）培养时间为15 d (20℃)，浓度为2 × 10^7~8孢子/mL，农杆菌-受体共培养温度为25℃，共培养时间为72 h。在最适条件下的转化效率达到80个转化子/百万分生孢子。T-DNA插入基因组的频率为100%，单拷贝插入频率为72.7%，转化子抗潮霉素性状能稳定遗传。从2136个转化子中获得了11个菌丝生长减缓突变体，7个色素产生缺陷突变体和14个分生孢子产生缺陷突变体，并从这些突变体中鉴定出7个致病力丧失突变体。采用hiTAIL-PCR技术，从3个突变体中获得了T-DNA插入位点侧翼序列。上述结果为深入研究L. biglobosa的生态适应性机制及致病机制提供了材料和线索。     

南瓜银叶突变体48a的表型特征及其遗传学分析

南瓜银叶突变体48a是在嫩食型中国南瓜中分离筛选到的稳定遗传自交系,银色叶不仅可以作为标记性状应用到育种中,还可为南瓜抗虫、抗病、耐寒等一系列研究提供重要的材料基础。笔者对银叶突变体的表型特征及叶片的解剖结构进行鉴定分析,发现植株整体长势、熟性与野生型无明显差异;成熟叶片正面全部呈银灰色,叶绿素含量明显降低,叶片上表皮细胞与栅栏细胞间明显剥离,存在明显的空隙。利用突变体48a和野生型49a南瓜自交系构建的六世代遗传群体,调查发现F2的绿叶与银叶符合3∶1的分离比,回交群体BC1P1分离比符合1∶1,表明南瓜银色叶性状由单隐性基因控制。     

玉米雄性不育K932MS不育株和可育株生理生化特性的比较研究

通过比较分析K932MS中不育及可育花药不同发育时期的可溶性糖、可溶性蛋白、游离脯氨酸含量及过氧化氢酶(CAT)、超氧化物歧化酶(SOD)和过氧化物酶(POD)活性差异,探讨其败育的生理生化机制。结果表明,不育花药中可溶性糖、可溶性蛋白含量在造孢细胞时期、花粉母细胞时期、二分体和四分体时期均显著低于可育花药,游离脯氨酸含量二分体和四分体时期显著低于可育花药;不育花药中CAT活性在造孢细胞时期显著高于可育花药,在花粉母细胞时期和四分体时期显著低于可育花药;SOD活性在花粉母细胞时期显著高于可育花药,在二分体和四分体时期都显著低于可育花药;POD酶活性在造孢细胞时期和四分体时期显著高于可育花药。造成K932MS败育的原因可能是由于不育花药中营养物质的严重不足,同时不育花药的CAT和SOD活性降低、POD活性升高,不利于花药的正常生长发育。     

基因编辑技术及其在作物育种中的应用与安全管理

基因组编辑技术是研究基因功能和对生物体基因进行定向改造的有力工具。随着近几年CRISPR/Cas9技术的快速发展,基因组编辑技术在作物育种领域起着越来越重要的作用。介绍了ZNFs、TALENs和CRISPR/Cas9系统的原理及在作物育种领域的研究进展,重点论述了CRISPR系统相关的变体和该系统在植物基因功能研究和作物育种中的进展。同时,也论述了基因编辑作物的检测方法及不同国家和地区对基因编辑作物的监管态度,重点介绍了美国、欧盟以及我国目前的监管态度,并分析了基因编辑作物存在的问题和发展趋势。为我国基因编辑作物的研究、安全管理和商业化批准提供了参考。     

Construction and Partial Biological Characteristics Analysis of Listeriolysin S llsB Gene Deletion Strain

In order to analyze the effect of listeriolysin S (LLS) llsB gene deletion on the biological characteristics of Listeria monocytogenes (LM),this study used homologous recombination to construct the llsB gene deletion strain LM90-ΔllsB,and the biological characteristics of growth characteristics,median lethal dose (LD₅₀) and organ-borne bacteria were studied in healthy Kunming male mice at 8 weeks old and weighing 40 g±5 g.The llsB gene deletion strain was successfully constructed,and the deletion strain had good genetic stability through continuous passage to 20 generations in vitro.Based on its growth curve examination,we found that the growth rate of the mutant strain was slightly higher than that of the parent strain.The results of mice infection test showed that the LD₅₀ of the parent strain and the deletion strain were 10^6.17 and 10^6.50 CFU,respectively.Compared with the parent strain,the amount of bacteria load of the deletion strain in the liver and spleen of the mice was extremely significantly decreased (P<0.01),the results showed that the infection ability of the mutant strain on mice was obviously weakened.No Listeria monocytogenes was detected in the brain.The results suggested that llsB gene might have direct or indirect regulatory effect on some biological characteristics of LM90,and it would provide a theoretical basis for further understanding of the pathogenic mechanism of LLS,prevention and control of listeriosis.     

水稻叶片早衰突变体的农艺与生理性状研究进展

叶片早衰严重影响水稻产量与品质。研究水稻叶片早衰突变体的农艺及生理性状,有助于了解水稻叶片衰老进程和机理。本文简述了叶片衰老特征和衰老机制,重点综述了水稻叶片早衰突变体的类型、农艺与生理性状研究进展及延缓水稻叶片衰老的栽培调控途径。可为水稻抗衰老品种选育及栽培调控提供理论依据。     

The mutagenesis of a type IV secretion system locus of Piscirickettsia salmonis leads to the attenuation of the pathogen in Atlantic salmon,Salmo salar

Piscirickettsiosis is a threatening infectious disease for the salmon industry, due to it being responsible for significant economic losses. The control of outbreaks also poses considerable environmental challenges. Despite Piscirickettsia salmonis having been discovered as the aetiological agent of the disease more than 25 years ago, its pathogenicity remains poorly understood. Among virulence factors identified so far, type four secretion systems (T4SS) seem to play a key role during the infection caused by the bacterium. We report here the genetic manipulation of P. salmonis by means of the transference of plasmid DNA in mating assays. An insertion cassette was engineered for targeting the icmB gene, which encodes a putative T4SS‐ATPase and is carried by one of the chromosomal T4SS clusters found within the genome of P. salmonis PM15972A1, a virulent representative of the EM‐90‐like strain. The molecular characterization of the resulting mutant strain demonstrated that the insertion interrupted the target gene. Further in vitro testing of the icmB mutant showed a dramatic drop in infectivity as tested in CHSE‐214 cells, which is in agreement with its attenuated behaviour observed in vivo. Altogether, our results demonstrate that, similar to other facultative intracellular pathogens, P. salmonis’ virulence relies on an intact T4SS.     

文心兰新品种‘海文吉祥1号’

文心兰新品种‘海文吉祥1号’是由文心兰‘柠檬绿’（Honey Angle）组培苗芽变株选育而来。花梗直立坚挺，花瓣和萼瓣淡黄色，唇瓣柱头处呈黄色，向四周边缘逐渐变浅成乳白色。为切花型品种，观赏价值高，应用前景广。     

高留胚糯稻突变体原胚糯的选育与营养成分分析

为创新高营养糯稻品种,采用250 Gy60Co-γ射线诱变处理宁粳3号种子,结合组织培养技术,选育出留胚率高达80%以上的优质高产抗病糯稻突变体原胚糯;同时,对原胚糯留胚米、宁粳3号糙米和精米进行蛋白质、矿物质、氨基酸等营养成分的测定,并对三者进行比较分析。结果表明,原胚糯留胚米蛋白质含量达8.25%,主要矿物质K、Ca、Mg、Na、Gu、Zn、Fe含量分别为898.41、107.55、95.13、21.39、1.11、12.12、10.51 mg·kg~(-1),17种氨基酸总含量达117.2 g·kg~(-1),人体必需氨基酸含量达39.6g·kg~(-1),均显著高于宁粳3号精米,具有与宁粳3号糙米相当的营养价值。本研究结果为进一步开发和利用原胚糯奠定了基础。