水稻抽穗期基因OsDof6功能的初步研究

【目的】以前期通过穗发育芯片筛选到一个水稻Dof家族转录因子OsDof6为研究对象,进一步探究OsDof6的表达模式与生物学功能。【方法】对OsDof6的基因、蛋白及启动子序列进行生物信息学分析,利用CRISPR/Cas9技术对该基因进行定点编辑,通过实时定量PCR和亚细胞定位技术分析该基因的表达模式。【结果】对该基因的启动子序列分析表明,该基因启动子中存在大量与光响应、激素响应及胁迫响应相关的顺式调控元件。利用CRISPR/Cas9技术获得了2种不同突变类型的功能缺失突变体9522^Dof6-3和9522^Dof6-4。对突变体T₁株系进行表型观察发现,相较于对照,两种突变体在营养生长阶段分蘖数明显降低,转入生殖生长阶段后,两种突变体的抽穗时间推迟约3 d。实时定量PCR结果显示OsDof6在水稻根、茎、叶和穗中均有不同程度的表达,在穗发育后期相对表达量明显提高;通过亚细胞定位分析发现OsDof6定位于细胞核。【结论】初步判断OsDof6基因会影响水稻分蘖数与抽穗期。     

Use of two PGPR strains in the integrated management of blast disease in rice (Oryza sativa) in Southern Spain

Biocontrol capacity of two plant growth-promoting rhizobacteria (PGPR) strains, against blast disease in rice paddy fields in Southern Spain was studied in three cropping seasons. Both strains (Pseudomonas fluorescens Aur 6 and Chryseobacterium balustinum Aur 9) had already shown biocontrol capacity against pathogens, ability to induce systemic resistance against leaf pathogens and against salt stress in different plant species. Bacterial treatments were carried out on seeds and/or on leaves. Strains were inoculated individually and in combination. Protection against natural disease incidence was evaluated, and rice production and quality measured in 2005 and 2006 trials. In 2004, natural disease incidence was low (between 0.1% and 0.35% of damaged leaf surface) due to environmental conditions; under these conditions, both strains significantly protected plants against rice blast. In 2005, disease incidence was higher than in 2004, reaching higher values of affected leaf surface in controls. In these conditions, each strain individually protected rice against rice blast, although the combination of both strains was the most effective treatment. All three treatments (Aur 6, Aur 9 and Aur 6 + Aur 9) reached 50% protection in panicles, with Aur 9 being the most effective. In 2006, the most effective treatment was the combination of both strains on leaves in three physiological stages, suggesting a biocontrol mediated protection. On the other hand, when bacteria were applied to seeds, disease incidence decreased up to 50%, suggesting induction of systemic resistance. Finally, a direct relation between protection mediated by the PGPR and the increase in rice productivity (mT/ha) and quality (weight of 1000 seeds and number of intact grains after milling) was found.     

淡紫拟青霉E7菌株固体放大发酵产孢培养基和工艺条件研究

为探讨淡紫拟青霉E7菌株固体放大发酵因子间的关系以获得大量孢子用于田间试验,通过正交试验设计优化固体发酵培养基成分和培养条件组合,并测定外加碳源、氮源、无机盐和装料方式对产孢量的影响。结果表明：以玉米粉+甘蔗渣+麸皮+壳聚糖组成的正交2号配方为最佳复合培养基质,分生孢子产量达7.13×109个/g,以发酵液pH+液相发酵终点+温度+初始含水量组成的正交4号配方为优适培养条件,分生孢子产量达8.97×109个/g;该菌株在优化后的培养基配方中添加0.4％蔗糖、0.2％蛋白胨、0.002％硫酸锰,以双层纱布袋装料按优化后的培养条件放大培养,产孢量最高可达到8.22×1010个/g。优化后的E7菌株发酵产孢培养基基质用量少,发酵成本低,适合于固体放大发酵生产淡紫拟青霉孢子。     

优质花生新品种唐花9号的选育及应用前景分析

唐花9号花生新品种,是唐山市农业科学研究院采用生物技术与常规技术相结合的方法,将优质与高产相结合,经过多年的单株定向选择,培育出的适合市场需求、符合外贸出口标准的花生新品种.2006年通过河北省鉴定.     

史氏鲟精子超微结构

利用扫描和透射电镜观察了史氏鲟精子形态和超微结构。精子具有顶体、头部、中段和单鞭毛等部分。顶体长0.99±0.06μm,宽0.87±0.09μm。细胞核从后往前逐渐变细,前端宽度为0.88±0.04μm,后端宽度为1.26±0.06μm,细胞核长7.29±0.32μm。核内含有3条核管(E),核管上行至顶体下行至植入窝而止。中段紧接头后部,长为0.51±0.12μm,宽0.91±0.05μm。中段含有线粒体、中心粒复合体和鞭毛的起始部分。线粒体分2~3层排列,线粒体中可见髓样嵴结构。在细胞核与中段接合部细胞核向内凹陷形成植入窝,纤维体位于植入窝内,其后是中心粒复合体。中段后缘延长为袖套,袖套腔中含有线粒体、脂质空泡。鞭毛从袖套中伸出,由轴丝组成,为典型的"9+2"微管结构。     

圆斑星鲽sox9基因的克隆与表达

本研究筛选到圆斑星鲽(Verasper variegatus)的性别相关基因sox9,并通过RACE技术获得了全长序列,基因全长为3287 bp,包括1431 bp的ORF,编码477个氨基酸,368 bp的5¢ UTR和1488 bp的3¢ UTR。在3¢ UTR中有多聚腺苷酸尾和加尾信号AATAAA。通过荧光定量PCR测定了sox9基因在圆斑星鲽成鱼不同组织中的表达水平,发现sox9基因在圆斑星鲽的脑、眼、鳃、心、肝、胆、肠、精巢、卵巢、肾和肌肉等各个组织中都有不同程度的表达。在鳃、脑和精巢组织中检测到较高水平的sox9转录,其中精巢中的转录水平显著高于其他组织,sox9基因在性腺中的表达显示出性别两相性差异。其在精巢中的表达水平要显著高于卵巢,说明sox9基因与雄性性腺发育相关。通过测定sox9基因在圆斑星鲽幼鱼不同发育时期(20、30、40、50、60、70和80日龄)的表达水平,发现其在20~50日龄表达量逐渐下降,在60日龄时表达量上升,推测表达量上升可能与幼鱼性腺分化相关。     

Using Bacillus subtilis E20‐fermented soybean meal as replacement for fish meal in the diet of orange‐spotted grouper (Epinephelus coioides,Hamilton)

The potential of Bacillus subtilis E20‐fermented soybean meal (FSBM) as a partial alternative component of fish meal (FM) in fed diets of orange‐spotted grouper (Epinephelus coioides) was evaluated in this study. An FM‐based diet and seven diets containing 10%, 20% and 30% and 10%, 20%, 30% and 40% of FM replaced by soybean meal (SBM) and FSBM, respectively, were fed to grouper for 84 days to evaluate possible substitution levels of FM by tracking growth performance, digestive enzyme activity and morphological changes in the liver and distal intestine. No significant differences in survival and muscle composition of grouper were found between controls and treatments. Growth performance and feed efficiency of fish fed diets with FM replaced by FSBM up to 30% were not significantly different from controls, whereas significantly decreased growth performance and feed efficiency occurred with diets containing >20% of SBM. Based on the feed efficiency, the maximum substituted levels of FM by SBM and FSBM in grouper diets were 18.36% and 29.32%, respectively, based on broken‐line analyses. Histopathological changes in the liver and distal intestine, and significantly lower activity levels of digestive enzymes, including pepsin in the stomach and trypsin, chymotrypsin, amylase and lipase in the distal intestine, were found in fish fed a diet containing 30% of FM replaced by SBM. However, these parameters were improved by the substitution of FSBM. It is therefore believed that FSBM has great potential to be used as a protein source in grouper diets in partial replacement of FM.     

不同维生素E水平饲料对云纹石斑鱼幼鱼低盐胁迫前后抗氧化和渗透压调节功能的比较

维生素E(VE)在鱼类繁殖、生长代谢、抗氧化能力和免疫机能等方面有至关重要的作用。为探究饲料VE添加量对云纹石斑鱼低盐度胁迫下抗氧化和渗透压调节功能的作用,本实验设计0、20、40、80和160 mg/kg等5个VE水平饵料添加量,依次为A、B、C、D、E组,饲喂云纹石斑鱼幼鱼56 d。然后进行6 h低盐度(12)胁迫实验,测试皮质醇(COR)、葡萄糖(GLU)、乳酸(LD)等血清生化指标,超氧化物岐化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-Px)、总抗氧化能力(T-AOC)和丙二醛(MDA)等抗氧化指标,以及鳃Na⁺/K⁺-ATP酶(NKA)、Ca²⁺-ATP酶活力等。结果显示,血清COR含量最高为盐度胁迫后A(32.74±1.53) ng/mL,最低为胁迫前E(19.06±3.88) ng/mL;GLU含量最高为盐度胁迫后A(16.46±0.99) mmol/L,最低为养殖前O(7.90±0.34) mmol/L;LD含量最高为低盐度胁迫前C(1.94±0.15) mmol/L,最低为胁迫后E组(1.30±0.06) mmol/L。说明VE的添加可以增强云纹石斑鱼抗应激的能力,且在47.524~134.566 mg/kg(B~D)的添加范围,效果较好。血清SOD、CAT、GSH-Px活力、T-AOC和MDA含量与投喂饲料中VE水平密切相关,SOD最高为胁迫前C(105.29±9.07) U/mL,最低为胁迫后A(11.23±2.30) U/mL;CAT最高为养殖前O(4.09±0.17) U/mL,最低为胁迫后A(0.35±0.10) U/mL;GSH-Px最高为胁迫前E(972.58±55.35) U,最低为胁迫后A(47.90±10.64) U;MDA最低为胁迫前D(33.48±2.34) nmol/mL,最高为胁迫后A(101.79±7.79) nmol/mL。饲料中添加91.378~178.924 mg/kg(C~E)水平VE对其抗氧化能力有增强作用。VE可增强NKA活力,当VE添加量为91.378 mg/kg(C)时,低盐度胁迫对NKA影响较小。而添加91.378~178.924 mg/kg(C~E)水平VE可增强云纹石斑鱼Ca²⁺-ATP酶活力。研究表明,饲料中添加91.378~134.566 mg/kg范围的维生素E,可以增强云纹石斑鱼的机体活力、抗应激及环境胁迫能力。     

饲料中添加不同水平的维生素E对半滑舌鳎(Cynoglossus semilaevis)亲鱼繁殖性能及后代质量的影响

在饲料中分别添加0、200、1200 mg/kg的维生素E(DL-α-生育酚乙酸脂形式)配制成Ec、El和Eh三种试验饲料,饲喂3龄半滑舌鳎亲鱼(雌性1.58±0.19 kg/尾、雄性0.20±0.05 kg/尾)70 d,探讨饲料中添加不同水平的维生素E对半滑舌鳎亲鱼繁殖性能的影响。结果显示,饲料中添加1200 mg/kg的维生素E(Eh组)不仅可以促进亲鱼性腺的发育,提高亲鱼产卵量、浮卵率、受精率、孵化率以及初孵仔鱼长度,而且影响仔鱼质量,仔鱼的成活率、12 d仔鱼长度、生存活力等方面均要高于对照组。而饲料中添加200 mg/kg的维生素E(El组)只对亲鱼性腺的发育和初孵仔鱼长度有一定的促进作用,对亲鱼产卵量、浮卵率、受精率、孵化率没有显著影响(P>0.05)。随饲料中维生素E的增加,血清中SOD的活性显著升高(P<0.05),MDA含量则显著下降(P<0.05)。此外,El和Eh试验组卵中干物质、维生素C的含量,以及卵子脂肪酸中PUFA、n-3PUFA、n-3HUFA和n-6PUFA的含量均要高于Ec组,El和Eh试验组之间的没有显著性差异(P>0.05)。结果表明,在半滑舌鳎亲鱼饲料中添加适量维生素E会亲鱼促进性腺的发育,增加产卵量,提高卵子和仔鱼的质量。在本试验中Eh组(1200 mg/kg)亲鱼的繁殖性能要优于Ec组(0 mg/kg)和El组(200 mg/kg)。     

用于生物质酶解过程的纤维素酶研究进展

纤维素酶解效率是木质纤维素经济、高效生化转化的限制瓶颈。该文讨论了影响纤维素酶酶解经济性与高效性的多个要素,如:高滤纸酶活菌株的选育、发酵、酶解机理、酶解影响因素及酶解混合体系的优化等。该文研究表明,纤维二糖水解酶可能是决定发酵体系中滤纸酶活高低的关键单酶组分,同时,该酶可能也是预处理后生物质酶解体系中决定滤纸酶活效率的关键单酶组分。酶解过程中关键限速反应的认识及关键限制因素形成机制的揭示将成为纤维素酶生化转化研究的重点,这些机理机制的建立可为构建高比活力纤维素酶提供理论依据。