地衣芽胞杆菌HS10原生质电转化体系的研究

地衣芽胞杆菌Bacillus licheniformis HS10及其粗蛋白对多种病原真菌具有防治效果,为挖掘其生防功能基因,本研究通过对原生质体的制备、再生培养基、渗透压稳定剂、电击参数等试验条件进行优化,建立了地衣芽胞杆菌HS10原生质体电转化法。结果表明最佳转化条件为:转接培养10h达到对数生长后期的菌体,浓缩4倍后,利用终浓度1mg/mL的溶菌酶,于37℃、150r/min酶解20min,酶解效率达到99.34%,1×SMM洗涤3次,调整菌浓度至1.0×108 cfu/mL,并通过1.6kV、5ms的电击后,迅速在SMMP溶液中100r/min、37℃恢复6~10h,涂布于含Kan和Erm的再生培养基DM3平板。将该方法用于突变体库构建的质粒pMarA(8 253bp)和基因敲除质粒pMADΔCP(13 043bp)的大片段质粒转化,分别获得了37cfu/μg DNA和10cfu/μg DNA阳性转化子。该遗传转化体系为地衣芽胞杆菌HS10中相关基因功能的研究提供了技术支撑。     

EMS诱变NC 55突变体库的创建及高钾突变材料的筛选

为了获得不同类型的烟草突变体库,用0.6%的EMS诱变处理NC 55,对诱变后的M 1代的大田性状进行调查分析,并从中筛选高钾突变材料。EMSNC 55 M 1代植物学性状和农艺性状变异类型丰富,突变率为11.88%,主要为多头突变、株型突变、叶片突变等;通过大田筛选和烟叶化验,筛选到钾含量高于对照NC 55的突变材料74份,其中钾含量超过1.50%的35份,以EMSNC 55-5501烟叶钾含量最高,为2.03%。EMSNC 55 M 1代单株留种606份,为进一步筛选各种类型突变体和选育新品种创建了很好的突变体库。     

Effects of fish meal replaced by fermented soybean meal on growth performance,intestinal histology and microbiota of largemouth bass (Micropterus salmoides)

This study was conducted to investigate the effects of replacing fish meal (FM) with fermented soybean meal (FSM) and soybean meal (SM) on growth performance, intestinal histology and microbiota of largemouth bass (Micropterus salmoides). The basal diet contained 350 g/kg FM (CON), and then, FM was replaced with SM and FSM at the ratios of 30% and 60% (SM‐30, SM‐60, FSM‐30 and FSM‐60), respectively. The largemouth bass (4.43 ± 0.13 g) were fed for 8 weeks. The results showed that weight gain of fish fed with FSM‐60 and SM‐60 diets was significantly lower, and feed conversion ratio of SM‐30, SM‐60 and FSM‐60 groups was significantly higher than the CON group (p < .05). The intestinal villus height of SM‐60 group and the villus width of SM‐60 and FSM‐60 group were significantly lower than the CON group (p < .05). The 30% FM replacement by SM and FSM significantly increased the abundance of Cetobacterium and Mycoplasma, respectively (p < .05). In conclusion, FSM could replace 30% FM in diet without negative impacts on the growth performance of largemouth bass, while the SM should be controlled below 30%.     

功能性食品黄精山楂酸奶的配方筛选

目的 探索一款以黄精、山楂、牛奶为主要原料的酸奶配方。方法 通过正交设计及感觉评分研究白糖、奶粉、黄精药液、山楂药液对酸奶成品色、香、味、组织状态的影响。结果 黄精山楂酸奶以奶粉180 g/L，山楂药液（0.6 g生药/mL）50 mL/L，黄精药液（0.6 g生药/mL）100 mL/L，白糖80 g/L作为配方，酸奶成品色泽、口感、风味较佳。质量分析显示黄精山楂酸奶脂肪含量3.2%，非脂乳固体8.3%，蛋白质3.2%，酸度78°T，乳酸菌群1.3×10⁶ CFU/mL，大肠菌群5 MPN/mL，致病菌（沙门氏菌、金黄色葡萄球菌）未检出。感官特性、微生物指标和理化指标都符合国家标准GB19302-2010《发酵乳》的相关规定。结论 黄精山楂酸奶的制备具有可行性，为中医药开发新型功能性食品提供新思路。     

茉莉酸合成抑制剂对棉花脱分化及茎尖培养的影响

以棉花幼苗下胚轴和茎尖作为试验材料,通过添加不同浓度的外源茉莉酸合成抑制剂,观察棉花下胚轴生根及愈伤组织的形成率以及对茎尖分化的影响,探究茉莉酸合成抑制剂对棉花脱分化及茎尖培养的影响。结果表明,茉莉酸合成抑制剂可以显著地促进愈伤组织的生成,加快生根,但是会在一定程度上抑制愈伤组织POD及SOD的活性。当茉莉酸合成抑制剂浓度为0.075 mg·L~(-1)时,对生根及愈伤组织形成方面作用效果最显著。而在茎尖培养过程中,2.0 mg·L~(-1)的茉莉酸合成抑制剂在棉花茎尖培养初期对分生组织分化起显著促进作用,棉花生长量最大,植株SOD酶和POD酶活性最大。     

Construction and evaluation of introgression lines and fine mapping of ehd8 from Jinghong common wild rice (Oryza rufipogon)

Heading date is one of the most important traits in rice and regulated by multiple genes. Common wild rice is the ancestor of Asian cultivated rice and harbours abundant genetic diversity. To use wild rice resource in rice breeding, a set of 154 introgression lines (ILs) covering 93% of the genome of Jinghong common wild rice was constructed in the background 'Teqing', using 208 simple sequence repeat markers evenly distributed on 12 chromosomes. Among the ILs, the line JIL64 displayed late heading independent of photoperiod. Genetic analysis using the two F₂ populations crossed ''Teqing'/JIL64 and JIL64/'Teqing' revealed that late flowering was controlled by a recessive gene on chromosome 8 (designated early heading date 8, ehd8), and ehd8 was fine mapped to the 50‐kb region flanked by markers RM22221 and 64Indel4. Sequencing and qRT‐PCR demonstrated that LOC_Os08g01410 and LOC_Os08g01420 were deleted in JIL64 and may be associated with the late heading of Jinghong common wild rice. These findings lay a practical foundation for characterizing ehd8, and the ILs help to mine genes from Jinghong common wild rice.     

Suppression subtractive hybridization and microarray analysis reveal differentially expressed genes in the <Emphasis Type="Italic">Lr39/41</Emphasis>-mediated wheat resistance to <Emphasis Type="Italic">Puccinia triticina</Emphasis>

Wheat leaf rust caused by Puccinia triticina (Pt) is one of the most severe fungal diseases threatening the global wheat production. The use of leaf rust resistance (Lr) genes in wheat breeding programs is the major solution to solve this issue. Wheat isogenic line carrying the Lr39/41 gene has shown a moderate to high resistance to most of the Pt pathotypes detected in China. In the present study, a typical hypersensitive response (HR) was observed using microscopy in leaves of the Lr39/41 isogenic line inoculated with the avirulent Pt pathotype THTT from 48 h-post inoculation. Two Lr39/41 resistance-associated suppression subtractive hybridization (SSH) libraries with a total of 6000 clones were established. Microarray hybridizations were performed on all obtained SSH clones using RNAs extracted from leaves of the Pt-inoculated and non-inoculated Lr39/41 isogenic lines, and leaves of the Pt-inoculated and non-inoculated Thatcher susceptible lines. Differentially expressed clones were analyzed by significance analysis of microarrays (SAM), followed by further sequencing. A total of 36 Lr39/41-resistance-related differentially expressed genes (DEGs) were identified, many of which had been previously reported to be involved in the plant defense response. The expression levels of eight selected DEGs during different stages of the Lr39/41-mediated resistance were further quantified by a qRT-PCR assay. Several pathogenesis-related (PR) and HR-related genes seem to be crucial for the Lr39/41-mediated resistance. In general, a brief profile of DEGs associated with the Lr39/41-mediated wheat resistance to Pt was drafted.     

入侵杂草虉草在云南的发生、危害和防除状况调查

采用倒置"W"九点取样法和5级目测法,调查了云南冬季作物田杂草虉草的发生和危害状况。结果表明,截至2017年在云南有7个地州33个区县发现有虉草危害,其中保山地区危害最为严重,占全省受害面积的92%,其次是大理白族自治州。虉草在冬季作物田大麦、小麦、油菜和蚕豆田均有发生。危害的虉草有两个种,分别是小籽虉草Phalaris minor和奇异虉草P.paradoxa。其中小籽虉草的发生较为普遍,奇异虉草为零星发生。虉草造成的危害指数在6%~50%之间,对农作物造成的产量损失在40~110kg/667m2之间。虉草的防除以化学防除为主,人工防除为辅,但目前尚缺乏经济有效的防除农田虉草的除草剂。     

高邮湖大银鱼、太湖新银鱼Cytb和COⅠ 基因序列多态性分析

为探明高邮湖大银鱼、太湖新银鱼野生资源状况,利用线粒体DNA Cytb和COⅠ基因序列,对高邮湖大银鱼、太湖新银鱼的遗传多样性水平及遗传结构进行分析。试验结果显示,大银鱼Cytb基因序列全长1141 bp,其中多态性位点14个,共定义12个单倍型,单倍型多样性和核苷酸多样性分别为0.871±0.031和0.00172±0.00019,具有高单倍型多样性和低核苷酸多样性特征。COⅠ基因片段长度为630 bp,其中多态位点5个,共定义6个单倍型,单倍型多样性和核苷酸多样性分别为0.747±0.041和0.00202±0.00019,具有高单倍型多样性和低核苷酸多样性特征;太湖新银鱼Cytb基因序列全长1141 bp,其中多态性位点13个,共定义9个单倍型,单倍型多样性和核苷酸多样性分别为0.609±0.078和0.00094±0.00027,具有高单倍型多样性和低核苷酸多样性特征。COⅠ基因片段长度为630 bp,其中多态位点2个,共定义3个单倍型,单倍型多样性和核苷酸多样性分别为0.232±0.085和0.00038±0.00014,呈现低单倍型多样性和低核苷酸多样性。大银鱼和太湖新银鱼Tajima′s D和Fu′Fs中性检验值为负值,且歧点分布曲线呈单峰型,表明历史上经历过种群扩张。研究结果表明,应通过多种措施加强高邮湖银鱼种质资源保护。     

Comparative study on the utilization of crystalline methionine and methionine hydroxy analogue calcium by Pacific white shrimp (Litopenaeus vannamei Boone)

The objective of this study was to compare the supplemental effects of crystalline DL‐methionine (DL‐Met) and methionine hydroxy analogue calcium (MHA‐Ca) on growth performance of Pacific white shrimp. Eight isoproteinic (355.3 ± 2.0 g/kg diet) and isolipidic (70.0 ± 2.2 g/kg diet) diets were prepared as positive diet (20% fish meal), negative diet (15% fish meal) and DL‐Met, MHA‐Ca‐supplemented diets with the supplementation of 0.03%, 0.06%, 0.09% DL‐Met and 0.04%, 0.07%, 0.1% MHA‐Ca in negative diet respectively. Pacific white shrimp (0.92 ± 0.03 g) were fed one of the eight diets for 49 days. The results showed that dietary DL‐Met did not affect weight gain (WG) and feed conversion ratio (FCR) (p > 0.05), but the supplementation of 0.1% MHA‐Ca significantly increased WG, protein and lipid retention, and reduced FCR (p < 0.05) when compared to the negative group, and reached the similar levels as the positive control. The total free amino acids (TFAA) in haemolymph of MHA‐Ca groups and PC, NC group peaked at the 3rd hr after feeding, but the peaking time of DL‐Met groups was advanced to the 2nd hr. Activities of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in haemolymph of all MHA‐Ca groups peaked at the 2nd hr, but DL‐Met groups peaked at the 1st hr after feeding. The above results indicated that the supplementation of 0.1% MHA‐Ca in a low fish meal diet could improve the growth performance and feed utilization of Pacific white shrimp, but dietary DL‐Met did not significantly affect the growth.