油茶织蛾生物学特性研究

Casmara patrona is one of the main borer pests damaging Camellia oleifera in China. Few reports about this insect were published. The life history, biological characteristics and natural enemies of C. patrona were investigated through field survey and laboratory observation in this study. The results showed that C. patrona had one generation in one year or two years, and overwintered as 3rd-5th instar larva in Zhejiang Province. About 88 percent of larvae began to pupate in early May, and the rest kept feeding till the next year. The adults could be observed from early June to early July, and the peak of emergence occurred in late June. The adults began to oviposit in late June and terminated in early July. The eggs started hatching in early July, and then the larvae bored into the branch to feed. The entrance holes of the larvae distributed predominately in the middle of the eastern and southern part of C. oleifera tree. The larva had five instars, and there were 10-50 days when the larva did not feed between consecutive instars.     

RNAi-mediated knockdown of INHBB increases apoptosis and inhibits steroidogenesis in mouse granulosa cells

Inhibins are members of the TGFβ superfamily and act as suppressors of follicle stimulating hormone (FSH) secretion from pituitary glands via a negative feedback mechanism to regulate folliculogenesis. In this study, the INHBB gene was knocked down by three RNAi-Ready pSIREN-RetroQ-ZsGreen vector- mediated recombinant plasmids to explore the effects of INHBB silencing on granulosa cell (GC) cell cycle, apoptosis and steroid production in vitro. Quantitative real-time polymerase chain reaction, Western blot, flow cytometry and ELISA were performed to evaluate the role of INHBB in the mouse GC cell cycle, apoptosis and steroid production in vitro. The results showed that the relative mRNA and protein expression of INHBB in mouse GCs can be significantly reduced by RNAi with pshRNA-B1, pshRNA-B2 and pshRNA-B3 plasmids, with pshRNA-B3 having the best knockdown efficiency. Downregulation of the expression of INHBB significantly arrests cells in the G1 phase of the cell cycle and increases the apoptosis rate in GCs. This was further confirmed by downregulation of the protein expressions of Cyclin D1, Cyclin E and Bcl2, while the protein expression of Bax was upregulated. In addition, specific downregulation of INHBB markedly decreased the concentration of estradiol and progesterone, which was further validated by the decrease in the mRNA levels of CYP19A1and CYP11A1. These findings suggest that inhibin βB is important in the regulation of apoptosis and cell cycle progression in granulosa cells. Furthermore, the inhibin βB subunit has a role in the regulation of steroid hormone biosynthesis. Evidence is accumulating to support the concept that inhibin βB is physiologically essential for early folliculogenesis in the mouse.     

The Expression of Long Noncoding RNA (H19) in Mouse Skeletal Muscle Development

This experiment was intend to study the changes of long non-coding RNA (H19) expression levels in skeletal muscle development and regeneration,and lay the foundation of its mechanism reach in skeletal muscle development.C2C12 cell line and ICR mice were used as experimental material,bioinformatics assay was used to exploit its non-coding character and low conservatism in different species,and the expressions of H19 in C2C12 cell differentiation,skeletal muscle development and the phase of muscle regeneration were detected by quantitative Real-time PCR (qRT-PCR).The results showed that,H19 expression levels in postnatal mouse skeletal muscle decreased with increasing age;during C2C12 cell differentiation,H19 mRNA increased gradually,and then maintained a high level;The expression of H19 was maintained at a high level through days 4 to 6 after injury.In consideration of its express character in C2C12 cell differentiation and skeletal muscle damage repair model,H19 may play an important role in promoting myogenesis and skeletal muscle regeneration.     

加工番茄系统抗性诱导促生菌的筛选鉴定及其促生防病效果

【目的】 筛选具有诱导番茄系统抗性的促生菌并验证其对加工番茄早疫病、灰霉病的防效,为新疆加工番茄病害的生物防治提供理论依据和菌种资源。【方法】 采用稀释涂布法,分离番茄等作物的根部土壤细菌,以无菌生理盐水和稀释至10⁴倍数未接菌的NB培养基为空白对照,利用发芽实验初筛加工番茄促生菌;用菌株发酵液灌根处理加工番茄幼苗,以清水处理为空白对照,测量叶片中茉莉酸含量,复筛系统抗性诱导菌株,分别挑战接种番茄早疫病、灰霉病病原菌,统计疫情指数和诱抗效果。测定菌株16S rDNA序列,初步进行菌种鉴定。【结果】 共分离到147株细菌,筛选得到19株显著(P<0.05)促进加工番茄种子萌发、增加根长的促生菌,有4株促生菌显著(P<0.05)提高加工番茄叶片中茉莉酸含量,防病验证最终得到3株能同时有效防治加工番茄早疫病、灰霉病的菌株,诱抗防效在27.59%~39.44%。菌株FY10、FY12、FY93鉴定为Bacillus atrophaeus,Pseudomonas wadenswilerensis和Bacillus pumilus。【结论】 获得3株具有系统抗性诱导功能且有效防治加工番茄早疫病、灰霉病的促生菌。     

美国贸易政策不确定性对中国畜产品价格时变冲击效应研究

本文基于2006年1月至2020年11月的中国猪、牛、羊等畜产品价格数据和美国贸易政策不确定性指数,利用带有随机波动率的时变参数向量自回归模型(TVP-SV-VAR模型),考察美国贸易政策不确定性对中国畜产品价格波动的影响。研究发现:美国贸易政策不确定性对不同畜产品价格波动的影响存在阶段性特征,会推动畜产品价格周期性变动。在欧洲债务危机时期、中美贸易蓬勃发展后期、特朗普政府上台初期以及中美贸易摩擦升级时期这四个时期,美国贸易政策不确定性对中国不同畜产品价格波动的影响表现出差异性特征。美国贸易政策不确定性对中国猪肉价格的影响程度最深,对牛肉和羊肉价格冲击程度最小。据此,提出根据美国贸易条件变动预期相机抉择,达到平抑畜产品市场变动的目的,并以猪肉市场稳定为关键,多措并举,发挥市场供求调节能力。     

浙江地区鹅星状病毒分离鉴定及其衣壳蛋白多克隆抗体的制备

鹅星状病毒(GAstV)是当前鹅养殖业的重要病原,易引起雏鹅内脏痛风症状和死亡,造成巨大经济损失。于浙江省内采集30份病料,进行核酸检测、病原分离和测序,并克隆其ORF2序列至pET-28a原核表达载体,转化BL21菌株后经诱导获得衣壳蛋白,免疫新西兰大白兔制备多克隆抗体。结果显示,临床死亡雏鹅剖检均发现典型内脏痛风症状,核酸检测鉴定为鹅星状病毒阳性,且出现不同基因型鹅星状病毒混合感染情况。共分离得到ZJC14和ZJLD20两个毒株,其中,ZJLD20在鹅胚和LMH细胞中均稳定增殖,但ZJC14并不能适应LMH细胞。病毒基因组测序显示,ZJC14与ZJLD20亲缘关系较远,ZJC14属于GAstV-Ⅰ基因型,而ZJLD20为GAstV-Ⅱ基因型。重组表达载体pET28a-ORF2诱导后获得纯化目的蛋白,经免疫成功制备兔源多克隆抗体,该抗体可与目的蛋白结合。此外,间接免疫荧光和Western-blot试验结果显示,ZJLD20衣壳蛋白制备的多克隆抗体可与病毒结合反应。研究成果有利于后续对该病原致病能力的研究,同时,试验制备的ORF2衣壳蛋白与多克隆抗体为鹅源星状病毒感染的诊断试剂开发奠定了基础。     

小麦施用生物炭下化肥减施潜力研究

利用砂姜黑土区小麦玉米轮作制下开展的连续3年定位试验,分析了生物炭与化肥配施对小麦产量、产量要素、氮素吸收量、氮素回收率及土壤物理性状的影响,以期为砂姜黑土区小麦生物炭施用下化肥合理减量施用提供依据.结果表明,施肥仍是砂姜黑土区小麦高产的关键.在施氮量为225 kg/hm2水平下,生物炭与化肥配施,减少10％的氮肥,小麦可以达到稳产以及略有增产的效果,氮肥减量达20％时,小麦产量略有下降,氮肥减量幅度达30％以上时,小麦产量显著下降.生物炭与化肥配施下减量施肥可以提高氮素回收率,随着试验年限增长,氮素回收率有增加趋势.与单施化肥相比,施用生物炭土壤容重下降0.04～0.06 g/cm3,降幅2.92％～4.38％,土壤田间持水量提高2.97～4.47百分点,增幅11.8％～17.8％,方差分析结果,差异均达显著水平.因此,在淮北砂姜黑土冬小麦种植上,配施生物炭30 t/hm2,较常规施肥减少10％的氮肥、20％的磷钾肥,可以保障小麦增产稳产,提高氮肥回收率.     

fl(2)d单等位基因的敲除显著降低小菜蛾的生殖力和育性

【目的】RNA甲基化是基因转录后水平表观修饰的主要形式,参与了众多重要的细胞学过程。小菜蛾（Plutella xylostella）是危害十字花科蔬菜的重要寡食性害虫,与RNA甲基化相关基因的功能尚未见报道。本研究通过克隆小菜蛾的RNA甲基化蛋白同源基因fl(2)d,鉴定其表达模式,并敲除该基因以探究其生物学功能。【方法】通过小菜蛾基因组网站查找fl(2)d基因序列,PCR扩增其蛋白质编码序列（CDS）;采用实时荧光定量PCR（qRT-PCR）技术,检测小菜蛾不同发育阶段个体以及成虫生殖腺中fl(2)d的相对表达量;运用CRISPR/Cas9结合卵的显微注射技术,对小菜蛾fl(2)d进行编辑;将fl(2)d被编辑过的成虫与野生型成虫杂交,并对其产生的后代进行近交,筛选fl(2)d突变品系;观测并比较突变体与野生型个体遗传特性、生物学参数和表型的差异,明确fl(2)d的功能。【结果】克隆得到长度为912 bp的fl(2)d CDS,fl(2)d在雌蛹、雌成虫和卵中的表达量较高,雄成虫和雄蛹的表达量较低,幼虫期的表达量最低,成虫卵巢中表达量显著高于精巢。通过向小菜蛾的卵注射靶向fl(2)d的向导RNA（sgRNA）和Cas9蛋白的混合物,对所产生的阳性后代进行10代的单对近交筛选,获得3种杂合的移码突变品系,分别缺失了4个（Δfl(2)d213-4）、5个（Δfl(2)d213-5）和7个（Δfl(2)d214-7）碱基。在上述品系的筛选过程中,发现了6只缺失4个碱基的纯合突变个体,2只缺失5个碱基的纯合突变个体;缺失4个碱基的纯合个体成功配成了两对,近交未产卵;剩余的2只缺失4个碱基的雄性纯合个体和2只缺失5个碱基的雄性纯合个体分别与同世代的雌性杂合突变个体近交后仍未产卵。说明fl(2)d纯合突变的个体存活率极低,且可能无法产生后代。通过分析后代基因型的分离比,发现杂合突变个体近交以及杂合突变个体与野生型个体杂交产生的后代中,杂合突变个体与野生型个体的比例分别略小于2和1,说明fl(2)d杂合突变会影响小菜蛾正常的生长发育,并导致部分个体死亡。杂合突变体后代中含有突变的雌雄个体比例接近1﹕1（P<0.05）,推测小菜蛾fl(2)d可能与性别决定无关。只要是有突变品系小菜蛾所参与的交配,雌成虫产卵量和卵的孵化率均显著低于野生型（P<0.01）,所产的卵多数发育异常,表现为失水皱缩、不能正常孵化。通过对成虫的生殖腺进行解剖,发现在野生型雌成虫与突变体雄成虫交配后,卵巢内卵的附着量较未交配的个体明显减少;未交配的突变体雌成虫卵巢内卵的附着量亦少于野生型,而突变体雄成虫的精巢未见明显异常。部分能够孵化的杂合突变个体在整个发育过程会发生不同程度的畸变,导致不能正常完成整个世代;另外一些杂合突变个体未见异常,可以将突变类型遗传给后代。根据上述发现,提出了基于fl(2)d的小菜蛾遗传防控模型。【结论】fl(2)d参与小菜蛾的生殖过程和胚胎发育,突变后显著影响后代种群数量,是开展小菜蛾遗传控制的理想靶标。     

合肥草莓基地土壤重金属污染风险评价

采集合肥市某草莓基地区域100个样点的土壤,测定表层土壤（深度分别为0～10 cm和10～20 cm）的6种重金属（Cd、Hg、As、Pb、Cr和Cu）,利用地累积指数方法对草莓种植土壤重金属的污染程度和生态风险进行评价,并结合多元统计分析探讨重金属来源。结果表明,Pb、Cd、Hg、Cu和Cr虽对草莓基地部分区域的土壤有一定程度的累积和污染,但污染风险低;主成分分析表明,Pb、Cd、As、Cu和Hg源于人类农业活动,Cr源于土壤母质及风化累积。显示草莓基地整体区域的土壤重金属生态风险低,种植基地土壤重金属未对草莓质量安全产生影响。     

外源ABA对非生物胁迫下黄芩抗逆性的影响研究

为探讨外源脱落酸(ABA)对非生物胁迫(干旱、低温、高温)下黄芩抗逆性的影响,以一年生商洛黄芩幼苗为试验材料,采用盆栽试验研究了不同浓度(10、25、50、100、150μmol/L)ABA对黄芩幼苗POD、SOD、CAT酶活性,以及质膜透性、叶绿素、MDA含量的影响.结果表明:ABA能有效地改善黄芩抗干旱、低温、高温胁迫能力,其中以喷施25μmol/L ABA效果最好,其在干旱情况下叶绿素含量、POD、SOD、CAT酶活性分别是干旱对照的1.33、1.69、1.13、1.40倍,MDA含量和相对电导率分别下降了9.29 nmol/g和3.62个百分点;其在低温情况下叶绿素含量比低温对照提高了1.2 mg/g,POD、SOD、CAT酶活性分别是低温对照的2.00、1.15、2.79倍,MDA含量和相对电导率分别下降了11.99 nmol/g和4.0个百分点;其在高温情况下叶绿素含量、POD、SOD、CAT酶活性分别是高温对照的1.40、1.44、1.09、1.46倍,MDA含量和相对电导率分别下降了10.06 nmol/g和3.1个百分点.由此可知,喷施25μmol/L ABA能明显改善黄芩苗的生理特性,为黄芩生产中产量、品质及药用价值的提高提供参考.