核盘菌Ss160的基因克隆与功能初探

由核盘菌（Sclerotinia sclerotiorum）引起的菌核病是油菜等十字花科作物的重要病害，对油菜的产量品质造成严重影响。本研究基于转录组数据筛选到一个核盘菌基因Ss160 (Sscle04g035160)，发现Ss160在接种油菜的核盘菌中高度表达。生物信息分析表明该基因具有信号肽，且序列在真菌界高度保守。亚细胞定位显示Ss160无特异性的定位，烟草叶片瞬时表达Ss160在核盘菌离体叶接种实验中病斑扩展显著小于表达空载体的和野生型烟草叶片，暗示异源表达Ss160能够提高植物的菌核病抗性。基于酵母系统的体外实验证明Ss160具有转录激活能力。该研究结果为后续Ss160在核盘菌-植物互作中功能机制的研究提供基础，也为油菜菌核病的研究提供借鉴和参考。     

Characterization of the petiole length in soybean compact architecture mutant M657 and the breeding of new lines

Phenotypic screening of soybean germplasm suitable for high planting density is currently the most viable strategy to increase yield. Previous studies have shown that soybean varieties with dwarf features and a short petiole often exhibit a compact plant architecture which could improve yield through increased planting density, although previously reported short petiole accessions were ultimately not usable for breeding in practice. Here, we established a method to assess petiole length and identified an elite mutant line, M657, that exhibits high photosynthetic efficiency. The agronomic traits of M657 were evaluated under field conditions, and appeared to be stable for short petiole across seven locations in northern, Huang–Huai, and southern China from 2017 to 2018. Compared with the Jihuang 13 wild type, the mutant M657 was shorter in both petiole length and plant height, exhibited lower total area of leaf, seed weight per plant and 100-seed weight, but had an increased number of effective branches and the growth period was prolonged by 2–7 days. Using M657 as a parental line for crosses with four other elite lines, we obtained four lines with desirable plant architecture and yield traits, thus demonstrating the feasibility of adopting M657 in breeding programs for soybean cultivars of high density and high yield.     

外源SLs和纳米K2MoO4对干旱胁迫下油菜种子萌发的影响

干旱胁迫是油菜种子萌发和生长发育的主要限制因子之一。以甘蓝型油菜品种Q2（抗旱型）和秦优8号（干旱敏感型）为试验材料,研究干旱对油菜种子萌发的影响,探讨外源生长调节物质独脚金内酯（SLs）和纳米材料（n-K₂MoO₄）对干旱胁迫下油菜种子萌发的生理调控作用。结果表明,干旱胁迫显著抑制种子萌发,SLs和n-K₂MoO₄均可显著提高发芽率,增加幼苗干重和子叶叶绿素含量,提高超氧化物歧化酶、过氧化物酶活性及可溶性糖、脯氨酸含量,降低丙二醛含量。0.10µmol/L GR24+0.24mmol/L K₂MoO₄对干旱胁迫下种子萌发和氧化损伤的缓解效应较好。外源SLs和n-K₂MoO₄通过提高油菜种子萌发期的保护酶活性和渗透调节物质含量来提高抗旱性。本研究表明植物激素和纳米材料在农业生产中具有应用价值。     

花生黄曲霉毒素产毒菌种群区系和地理分布

花生是我国重要的食油兼用作物，黄曲霉及其毒素分布广、危害大，可贯穿田间、储藏、加工、流通多环节，是威胁花生质量安全的主要风险因子之一。土壤黄曲霉菌和寄生曲霉菌是花生感染产毒的主要源头，系统研究花生田间黄曲霉菌和寄生曲霉菌的分布，对开展花生黄曲霉毒素污染预警和绿色防控意义重大。本文重点研究论述了花生田间黄曲霉菌和寄生曲霉菌分布的影响因素和种群生物地理分布特征等，提出了当前土壤黄曲霉菌和寄生曲霉菌分布研究中存在的问题和未来重点研究方向。     

HSA转基因水稻二重微滴数字PCR定量检测方法研究

重组人血清白蛋白（HSA, Human serum albumin）基因工程水稻是我国自主研发的可规模化生产水稻重组人血清白蛋白（OsrHSA）的转基因品系，具有极高的推广价值和应用前景，但是目前缺乏对其进行精准定量检测的方法。微滴数字PCR（ddPCR, droplet digital PCR）是近年来新兴的前沿PCR技术，不依赖标准物质即可实现对DNA分子的绝对定量，在转基因产品定量领域被广泛应用。本研究基于ddPCR平台建立了适用于重组人血清白蛋白基因工程水稻（114-7-2品系）的二重ddPCR精准定量检测方法。通过对引物探针的特异性进行验证、对引物探针工作浓度和反应退火温度进行优化，获得最佳反应条件。进而对该方法检测限、定量限和结果重复性做了测定。最后通过对不同含量的重组人血清白蛋白转基因水稻样品进行定量检测，分析比较传统实时荧光定量PCR（quantitative real-time PCR, qRT-PCR）和二重ddPCR的优劣，结果表明本研究建立的转基因水稻HSA/PLD二重ddPCR方法稳定性更好、灵敏度高、成本低廉，精准可靠，适用于不依赖标准物质的精准定量HSA转基因水稻转化体含量分析，可取代qRT-PCR方法进行HSA转基因水稻的绝对定量检测，完善了我国转基因水稻成分精确定量检测技术体系。     

Comparative advantages of chemical compositions of specific edible vegetable oils

As important supplementary to major edible oils, comparative chemical advantages of minor edible oils decide their development and usage. In this study, chemical composition of 13 kinds of specific edible vegetable oils were investigated. The comparative advantages of chemical compositions of these edible oils were obtained as follows: (1) camellia, tiger nut and almond oil were rich in oleic acid, the contents of which accounted for 79.43%, 69.16% and 66.26%, respectively; (2) safflower oil contained the highest content of linoleic acid (76.69%), followed by grape seed (66.85%) and walnut oil (57.30%); (3) perilla seed, siritch, peony seed and herbaceous peony seed oil were rich in α-linolenic acid (59.61%, 43.74%, 40.83% and 30.84%, respectively); (4) the total phytosterol contents of these oils ranged from 91.46 mg/100 ​g (camellia oil) to 506.46 mg/100 ​g (siritch oil); and (5) The best source of tocopherols was sacha inchi oil (122.74 mg/100 ​g), followed by perilla seed oil (55.89 mg/100 ​g), peony seed oil (53.73 mg/100 ​g) and herbaceous peony seed oil (47.17 mg/100 ​g). The comparative advantages of these specific edible oils indicated that they possess the high potential nutritional values and health care functions.     

通过CRISPR/Cas9技术突变BnMLO6基因提高甘蓝型油菜的抗病性

基因编辑技术可以实现对目标基因高效准确的修饰,为植物遗传改良开辟了新途径。霉菌抗性位点(Mildew resistance locus O, MLO)基因是植物对白粉病菌防御的主效负向调节因子,突变后能增强植物对白粉病的抗性,但在油菜中是否具有同样的功能尚未见报道。为解析该基因在油菜抗病中的功能,本研究通过分析油菜接种核盘菌后基因的表达情况发现,BnMLO6基因受核盘菌诱导表达;利用CRISPR/Cas9基因编辑技术获得了一份BnMLO6基因6个同源拷贝同时突变的材料mlo6-212。遗传分析表明,CRISPR/Cas9引起的BnMLO6基因突变能够稳定遗传;mlo6-212突变体在田间和温室条件下都表现出明显的白粉病抗性;在接种核盘菌24 h后,病斑面积显著低于野生型,减小19.5%;BnMLO6基因突变能激发叶片胼胝质的自发堆积,增强接菌后乙烯和茉莉酸抗病信号。因此,BnMLO6基因可能参与了多条抗病信号路径,负向调控油菜对白粉病和菌核病的抗性。研究结果不仅为BnMLO6基因协同调控油菜多种病原菌抗性的研究提供了参考,也为油菜抗病性遗传改良提供了抗性资源和技术支撑。     

膨爆预处理对三种菜籽油品质及挥发性风味成分的影响

本文以甘蓝型、白菜型和芥菜型三种油菜籽为原料，探究了油菜籽经过膨爆预处理后菜籽油的品质及挥发性风味成分的变化。研究结果表明，膨爆预处理后甘蓝型、白菜型和芥菜型三种菜籽油的酸值和过氧化值均显著增加（P＜0.05），分别增加了0.25、0.49、0.39 mg KOH/g和0.20、0.18、0.18 mmol/kg，均在国际食品法典规定的范围内；三种菜籽油的氧化诱导时间均显著增加（P＜0.05），分别增加了17.32、10.06和13.99 h。膨爆预处理对三种菜籽油的脂肪酸组成无显著影响。膨爆预处理后，甘蓝型、白菜型和芥菜型三种菜籽油中苯乙腈、3-甲基巴豆腈、5-己烯腈等含硫化合物含量、杂环类物质以及氧化挥发物的种类和含量均显著增加（P＜0.05），杂环类物质和氧化挥发物中分别是以吡嗪类和醛类物质为主。     

利用SSR标记研究甘蓝型油菜叶绿体基因组DNA多态性

叶绿体基因组具有母性遗传、单倍性、高度的保守性和明显的种内差异等特点。研究不同品种叶绿体基因多样化程度，可为甘蓝型油菜的收集与引种及遗传改良提供指导。利用15对特异性叶绿体SSR标记对287份国内外甘蓝型油菜叶绿体基因组多样性进行了分析，结果显示，在15对叶绿体SSR特异引物中，5对引物扩增产生多态性条带合计19条，平均每对引物检测出3.8条多态性条带；287份材料共划分成14个单倍型，优势单倍型H01占比74.91%、H02占比13.59%、H03占比4.88%，其余11种单倍型（H04-H14）占比合计6.62%；H02为波里马、陕2A胞质类型，为中国特有单倍型；国外甘蓝型油菜中，俄罗斯与瑞典的单倍型较丰富，引进俄罗斯与瑞典的资源能更高效拓宽中国甘蓝型油菜的遗传多样性本底。本研究还获得2对引物（MF-4和ccmp2）得到特异条带，并为此建立了快速鉴定波里马、陕2A胞质类型的方法，为波里马与陕2A细胞质类型材料的鉴定与保护提供了方法。