Construction of mouse ESC line stably expressing Pdx1 by Tet-On system

AIM To construct the mouse embryonic stem cell （ESC） line with stable pancreatic and duodenal homeobox 1 （Pdx1） expression by Tet-On system, which may lay a foundation for further research on the differentiation of Pdx1⁺ definitive endoderm cells into pancreatic cells. METHODS The Pdx1-overexpressing lentiviral vector with green fluorescent protein marker and puromycin resistance was constructed by Tet-On system and was used to infect the mouse ESC. The cells were divided into 3 groups： blank control group （ESC group）, empty lentivirus control group （PDX1^- ESC group） and Pdx1 lentivirus transfection group （PDX1⁺ ESC group）. Flow cytometry was used to detect the transfected cells after screening by doxycycline （DOX）. The function of Tet-On system and the expression of Pdx1 gene were detected. The transfected cells in PDX1^- ESC group and PDX1⁺ ESC group were sorted by flow cytometry, and constructed ESC line with stable expression of Pdx1 and negative control ESC line were verified. RESULTS （1） The positive rates of transfected cells in PDX1^- ESC group and PDX1⁺ ESC group were 90.72% and 94.01% after screening by DOX, respectively. The positive rates of transfected cells in PDX1^- ESC group and PDX1⁺ ESC group was 97.84% and 98.13% after sorting by flow cytometry, respectively. （2） With DOX, green fluorescence was observed in PDX1^- ESC group and PDX1⁺ ESC group. The mRNA and protein expression of Pdx1 was significantly increased in PDX1⁺ ESC group （P<0.05）. Without DOX, no green fluorescence was observed in the cells of the 3 groups, and no significant difference in the mRNA and protein expression of Pdx1 was observed （P>0.05）. （3） After 3 months of cryopreservation, the cell lines still survived in resuscitation culture and were regulated by DOX. CONCLUSION Using Tet-On system, the mouse ESC line with inducible Pdx1 expression were successfully established and could be used as an effective cell model to research the differentiation of Pdx1⁺ definitive endoderm cells into pancreatic cells.     

红叶石楠离体培养技术研究

以红叶石楠半木质化带芽茎段为外植体,探讨不同外源激素水平对其不定芽分化、增殖与不定根形成的影响。结果表明:MS BA 1.0~2.0 mg/L IBA 0.1~0.3 mg/L GA 0.5 mg/L的分化增殖效果好;增殖系数达6.5以上;White NAA0.1 mg/L AC(活性炭)0.5%或White IBA 0.1 mg/L AC 0.5%或White IAA 0.5 mg/L AC 0.5%的生根效果较好,生根率可达90.5%以上;在珍珠岩∶蛭石∶腐叶土=5∶2∶1的基质中炼苗,成活率可达96%以上。     

软枣猕猴桃‘奇异莓6号’离体再生技术

以软枣猕猴桃无芽茎段为外植体进行离体培养,具有成本低、取材方便、材料充足等优点,但以往的研究表明无芽茎段诱导再生率低。本试验以‘奇异莓6号’软枣猕猴桃无芽茎段为外植体,探究不同植物生长调节剂、光照强度、温度、苗龄对不定芽诱导的影响。结果表明：最佳外植体为苗龄30d的无芽茎段,在光照12h/d(光照强度50LX)、室温20℃下,不定芽诱导的最适培养基为MS+2mg/L ZT,不定芽以间接途径发生,培养40d无芽茎段两端开始形成淡绿色愈伤组织,培养45d愈伤组织表面出现红色小点,开始形成不定芽,培养55d愈伤组织诱导率、愈伤组织分化率及不定芽数均到最大,分别为100%、100%及18.87个/块。待苗长至3cm左右时转到MS+0.4mg/L ^IBA中进行生根培养,培养30d生根率可达100%、根数为8.53/株、根长1.68cm、根粗0.14cm。     

茎弯曲处理对番木瓜开花结果的影响

对“苏罗”和“台农2号”番木瓜2个品种植株进行茎弯曲处理,研究其对番木瓜开花结果的影响。结果表明,茎弯曲处理对“苏罗”和“台农2号”番木瓜都具有降低最低结果高度、增加长圆形果实和长圆形两性花数量的显著效应;当茎弯曲长度在20.00-35.00cm时,随着茎弯曲长度的增加,其对降低番木瓜最低结果高度、增加长圆形果实和长圆形两性花数量的效应愈强;茎弯曲处理对营养繁殖的“苏罗”和种子繁殖的“台农2号”番木瓜开花结果的影响趋势一致。     

油菜黑胫病菌和茎基溃疡病菌的LAMP检测方法的建立

油菜黑胫病和油菜茎基溃疡病分别由子囊菌Leptosphaeria biglobosa和L.maculans引起.我国油菜产区仅发现L.biglobosa,未发现L.maculans.因而,L.maculans是我国的对外检疫性对象.这两种真菌形态相似,引起的病害症状相似,给田间快速准确鉴定带来难度.本研究基于环介导等温...     

叶鞘对杂交水稻茎秆抗倒能力的影响研究

为了明确叶鞘对杂交水稻茎秆抗倒能力的影响,选取4个杂交水稻品种,研究了其基部节间包鞘及去鞘的抗折力差异和基部节间叶鞘的包茎层数、包茎厚度、存活数量及其与节间包鞘抗折力的相关性.结果表明,基部节间包鞘的抗折力是去鞘的1.4～1.9倍,去鞘后的抗折力较包鞘的降幅24.3％-46.8％,叶鞘在齐穗期对抗倒性弱的品种影响大,而...     

辣木的染色体制片优化及核型分析

以辣木分生组织为材料,采用酶解去壁低渗法制片,探讨不同取材部位、预处理方式和酶解时间对辣木染色体制片的影响,并对其进行核型分析,以期为辣木的起源、演化及遗传育种提供一定理论依据。结果表明:以辣木新枝茎尖为最佳取材部位,用饱和对二氯苯预处理2 h,再用4%纤维素酶和5%果胶酶混合物酶解4 h,制片所得染色体效果最佳。核型分析表明,辣木染色体属于小染色体,有28条,核型公式为2n=2x=2n=28m,属于1B类型,核型不对称系数60.29%,核型对称程度较高,这表明辣木在进化中可能处于比较原始类型。     

甜玉米茎秆强度相关性状的数量性状基因座定位

为选育抗倒伏玉米品种,挖掘甜玉米茎秆强度相关性状的数量性状基因座（quantitative trait locus,QTL）,应用复合区间作图法以甜玉米组合T49×T56的F₂为作图群体,通过测定F_2：3家系的茎秆穿刺强度、茎秆抗压强度和茎秆弯折性能3个性状进行相关性状的QTL定位。结果表明,遗传连锁图谱包含153个SSR标记位点,覆盖玉米基因组1 199.1 cM,平均图距7.83 cM。3个性状共检测到10个QTL,其中与茎秆穿刺强度相关的2个QTL位于第3、7染色体上,解释11.81%和22.15%的表型变异,与茎秆抗压强度相关的4个QTL位于第1、3、7染色体上,单个QTL可解释3.68%~33.26%的表型变异,与茎秆弯折性能相关的4个QTL位于第3、6、8染色体上,单个QTL可解释3.55%~18.58%的表型变异。第7染色体检测到1个同时控制茎秆穿刺强度和茎秆抗压强度2个性状的QTL,位于umc1015~umc1987标记区间,分别可解释11.81%和33.26%的表型变异,第3染色体检测到1个同时控制茎秆穿刺强度、茎秆抗压强度、茎秆弯折性能3个性状的QTL,位于umc1400~dupssr23标记区间,分别可解释22.15%、13.27%和18.58%的表型变异。3个茎秆强度性状共同检测到的主效QTL,可在育种实践中用于分子标记辅助选择和抗倒伏玉米品种的选育。     

茎用芥菜新品种1年2代繁育研究初报

为了探索福建省茎用芥菜1年2代繁育技术,选择6个茎用芥菜品种,分别进行冬春季正常繁育和夏秋季高山加代繁育的2代繁育试验,并对比各品种2代繁育的生育期、株高、株幅和种子产量。结果表明：所有参试茎用芥菜品种只要通过打破种子休眠、衔接好2代繁育的播种期都能完成1周年2代繁育,夏秋季高山加代繁育的全生育期比冬春季正常繁育平均缩短约50 d;冬春季繁育比夏秋季高山加代繁育株高增加31.2%～132.9%,株幅增加1.5%～21.8%,所有参试品种冬春季繁育的种子产量都极显著高于相应夏秋季高山加代繁育,种子产量增加17.8%～153.6%。     

Strategy to identify resistance to white mould associated with high yield for irrigated common bean in Brazil

Development of common bean cultivars with partial white mould resistance through breeding techniques has been a challenge in Brazil. As yet, lines/cultivars from breeding programmes have not been investigated for resistance; therefore, this study screened 107 lines/cultivars for their reactions to white mould in 14 preliminary trials conducted under irrigation. Thirteen resistant lines/cultivars (three of Andean origin) and six Mesoamerican cultivars (three intermediately resistant and three susceptible) were selected for further investigation. These lines/cultivars and the resistant control A195 were evaluated in six advanced trials and two straw tests to assess the effectiveness of the screening procedure. In 11 preliminary trials, screenings were performed under moderate/high or higher disease pressure. These pressures occurred in two advanced trials in which, when yields were averaged across moderate/high and high pressures, 10 Mesoamerican lines/cultivars selected for resistance yielded 14%, 23%, and 38% more than intermediately resistant cultivars, A195, and susceptible cultivars, with median disease ratings (1–9 scale) of 4.5, 5.7, 5.7, and 6.7, respectively. In the straw test, three Andean lines/cultivars (A195 included) and two susceptible cultivars in the field were among those with the highest levels of physiological resistance. Thus, field rating under high disease pressure and greenhouse rating did not correlate significantly, suggesting that field trials are critical to evaluating resistance and to identifying high-yielding beans. Therefore, lines/cultivars from breeding programmes assessed in field trials may provide a low cost and fast way to identify high-yielding bean cultivars with partial resistance to white mould in the subtropical southern hemisphere.