苹果‘长富2号’开花调控转录因子基因MdIDD7的克隆及表达分析

以‘长富2号’苹果为试验材料,采用RT-PCR的方法,从其芽中克隆得到INDETERMINATE DOMAIN(IDD)转录因子基因MdIDD7,其开放阅读框长度为1 626 bp,编码541个氨基酸。序列比对和结构域分析表明,该转录因子含有1个核定位信号和4个高度保守的锌指蛋白结构域。系统进化树分析表明,MdIDD7与白梨(Pyrus×bretschneideri,XP_009364602.1)、桃(Prunus persica,XP_007225628.1)和梅(Prunus mume,XP_008220893.1)聚在一起。实时荧光定量PCR表明,MdIDD7在‘长富2号’不同组织(茎、叶、花、果和芽)中均有表达,其中芽的表达量最高。花后40~60 d,MdIDD7在易成花品种‘烟富6号’芽中表达量显著高于难成花品种‘长富2号’。"小年"树顶芽组织中MdIDD7的表达量显著高于"大年"树。在花芽诱导前期,外源GA处理诱导MdIDD7下调表达,而蔗糖处理诱导其上调表达,说明MdIDD7响应激素和糖信号,促进苹果成花。     

Agronomic traits of soybean cultivars released in different decades after grafting record‐yield cultivar as rootstock

Breeders have seldom considered the selection for root traits during the genetic improvement in soybean. It is hypothesized that grain yield would be increased by the root function improvement, especially for the current cultivars. The objective of this grafting experiment was to determine the effect of record‐yield cultivars L14 or Z35 as rootstocks on agronomic traits of cultivars released in different decades. A total of 11 cultivars, released in different decades, were used to graft onto L14 or Z35 rootstocks. The agronomic traits were measured in the pot‐culture experiments. Grafting cultivars released in different decades onto L14 or Z35 rootstocks resulted in higher yield, 100‐seed mass and apparent harvest index as compared with those of non‐grafted or self‐grafted plants. Grain yield gain of cultivars grafted onto record‐yield cultivar rootstocks was 0.40 g/plant/year from 1966 to 2006, which was larger than that of non‐grafts and self‐grafts (0.27 g/plant/year). The yield of current cultivars should increase more if their root functions were improved.     

苹果赤霉素信号转导因子MdGAMYB的克隆和表达分析

以‘长富2号’苹果为试验材料,从其短枝顶芽中克隆得到1个赤霉素信号转导因子MdGAMYB,对其进行生物信息学和表达分析。结果表明,MdGAMYB的开放阅读框(ORF)长度为1 656bp,编码551个氨基酸,蛋白质分子量为59.741 kD。生物信息学分析表明MdGAMYB编码的蛋白存在多个糖基化位点和磷酸化位点;序列分析表明,Md GAYMB和其他物种的GAMYB蛋白有很高的相似性,均含有保守的R2R3 DNA结合域和GAMYB家族所特有的Box1,Box2和Box3保守区域;系统进化分析表明,Md GAYMB与梨、梅花、草莓、枣和葡萄等的GAMYB蛋白具有较高的同源性。实时荧光定量PCR分析表明,Md GAYMB具有组织表达特异性,在叶片、花和芽中的表达量较高。外源GA3处理抑制了花芽孕育和翌年成花,抑制MdGAMYB的表达。在易成花品种‘烟富6号’中的表达量高于难成花品种‘长富2号’。     

紫花地丁开放花与闭锁花的发育及可溶性糖与淀粉含量的研究

紫花地丁(Viola philippica)为典型的两型花自花受精植物,具有开放花和闭锁花混合繁育系统。通过对开放花和闭锁花花芽形态发育的比较发现:开放花与闭锁花在花芽发育早期形态相似,4轮花器官原基均正常发生。出现明显差异的时期为4轮花器官原基形成以后,小孢子发育时期为产孢细胞阶段,开放花的5个花瓣与5枚雄蕊继续发育,每个雄蕊有4个花药室;而闭锁花只有2枚雄蕊继续发育,每个雄蕊有2个花药室,其余雄蕊与所有的花瓣依然为器官原基状态,不再发育。通过对花芽与叶片可溶性糖与淀粉含量的检测发现,花器官原基形成之后,开放花与闭锁花形态出现明显差异阶段开始,随着花芽的发育,可溶性糖与淀粉含量均呈上升趋势,且开放花花芽中的含量均明显高于对应发育阶段闭锁花花芽;而开放花植株的叶片可溶性糖与淀粉含量均低于闭锁花植株叶片,说明开放花所需要的能量高于闭锁花,推测可溶性糖与淀粉含量的差异与两型花发育有一定的关系。     

三种融合标签对大熊猫轮状病毒结构蛋白VP6-VP7可溶性表达的影响

大熊猫轮状病毒(giant panda rotavirus,GPRV)是引起幼龄大熊猫腹泻的主要病原,对圈养大熊猫产生了较大的危害。轮状病毒结构蛋白VP6是一种载体蛋白,可介导黏膜免疫反应。VP7是轮状病毒结构蛋白中主要的中和抗原。因此,VP6-VP7的融合表达作为候选抗原对该病的防治具有重要的意义。传统大肠埃希菌原核表达存在表达量低、可溶性差以及纯度低等弊端。本研究使用醛缩酶(EDA)、谷胱甘肽S-转移酶(GST)、麦芽糖结合蛋白(MBP)3种融合标签,以实现获得表达量高和纯度高的GPRV-VP6-VP7重组表达蛋白。将扩增的VP6、VP7基因片段利用同源重组酶构建到含3种融合标签的表达载体pET21b上,将重组质粒转化至大肠埃希菌Rosetta(DE3)感受态细胞中进行低温诱导表达。用Ni-柱亲和层析法纯化目的蛋白,SDS-PAGE和Image J分析蛋白表达量和可溶性,Western blot分析得到表达的重组表达蛋白正确且具有蛋白活性。实验结果证明,EDA标签能显著促进VP6-VP7蛋白的原核可溶性表达,提高VP6-VP7蛋白表达量。     

甲状腺素对猪小肠上皮细胞miR-let-7a基因表达及细胞增殖的影响

miR-let-7a在动物细胞的分化、增殖与凋亡等方面发挥越来越重要的作用。甲状腺激素(TH)作用非常广泛,机体的每个细胞几乎都是TH作用的靶细胞,其可以促进组织分化、生长和成熟。本实验用甲状腺素(T4)浓度分别为(0、0.02、0.03、0.05、0.075、0.1、0.2μmol/L)在体外培养猪的小肠上皮细胞。结果表明:T4处理组的细胞体积形态相对于空白对照组没有明显变化;当T4添加浓度为0.03μmol/L时,细胞的增殖率显著低于其他组(P0.05);当T4浓度为0~0.03μmol/L时,let-7a的表达随着添加剂量的增加而升高,浓度从0.03~0.2μmol/L变化时,let-7a的表达呈现降低趋势,浓度为0.03μmol/L时表达量极显著高于其他组(P0.01)。let-7a的表达量与细胞增殖呈负相关。     

新疆与美国早实核桃(Juglans regia L.)品种生长结实与成花特性比较

以引进的美国核桃品种‘维纳’、‘艾米格’和新疆本地品种‘温185’和‘新新2号’为试材,研究分析不同品种的植物学和生物学特性、花芽分化过程以及相关成花基因的表达。结果显示:4个品种在树体生长、枝芽发育特性、物侯期以及成花率和坐果率方面没有明显差异。在结果枝同一坐果部位形成双果、三果或以上的能力‘新新2号’明显强于‘维纳’,但‘新新2号’的果实大小和单果重都低于‘维纳’,而‘维纳’平均结果母枝上形成侧花芽的数量和坐果率高于‘新新2号’。从萌芽到开花结束直至当年侧芽发育过程中‘维纳’混合芽中的JrFT和JrLFY基因表达也高于‘新新2号’,而同时期的JrFLC基因表达则相对较低,说明‘维纳’在花芽分化和花器官形成过程中通过高表达JrFT和JrLFY基因,低表达JrFLC基因促进结果母枝上形成大量侧花芽,并利用大量侧花芽抽生结果枝坐果,形成较大的果实,使得最终产量并不低于同一个坐果部位大量着生2个或多个果实的‘新新2号’品种。     

溪荪新品种‘芭蕾女’

 ‘芭蕾女’是从溪荪（Iris sanguinea）田间自然杂交实生后代中选育出的新品种。2年生植株盛花期株高60 cm 左右，冠幅近52 cm，每株丛花葶数18 ~ 25，单葶花2 ~ 3朵。花形独特优美，盛开时外花被平展，内花被垂直向上，花色为蓝紫色，花径约9 cm，连续花期 20 d 左右。可在哈尔滨地区露地越冬，适宜推广应用到北方地区栽培。     

不同叶龄蘖、穗氮肥组合对粳稻产量及氮素利用的影响

以主茎叶片数不同的粳稻品种吉粳88 (14片)、沈农265 (15片)和沈农1401 (16片)为试材,采用大田筒栽方式,在总施氮量225kghm–2及轻简施肥(基肥、蘖肥、穗肥)模式基础上,设置基蘖肥∶穗肥6∶4和8∶2两种施肥比例,并分设不同源、库叶龄期施氮组合即不同叶龄蘖、穗肥精确施氮组合。分析了不同源库期氮肥运筹模式对水稻农艺性状、产量及氮素利用特性的影响。结果表明:(1)在有效穗数、分化颖花数、产量和氮素利用率方面,吉粳88、沈农265、沈农1401不同氮肥运筹下最佳蘖、穗肥叶龄组合均为6∶4显著高于8∶2。(2)不同氮肥运筹下,吉粳88在8叶(叶龄指数57.1%)、沈农265在9叶(叶龄指数60.0%)、沈农1401在10叶(叶龄指数62.5%)时,即叶龄指数在60%左右时,施用蘖肥效果最佳,最终穗数最多,对保蘖起主要作用;吉粳88在11叶(叶龄指数78.6%)、沈农265在12叶(叶龄指数80.0%)、沈农1401在13叶(叶龄指数81.3%)时,即叶龄指数在80%左右时,施用穗肥效果最佳,最终穗粒数最多,对促花起主要作用。(3)吉粳88-6∶4 (8, 11),沈农265-6∶4 (9, 12),沈农1401-6∶4 (10, 13) 3组处理,在产量、氮素积累量、氮素吸收利用率、农学利用率及偏生产力等方面,显著高于同品种不同叶龄蘖、穗氮肥组合中的其他处理。因此,适当延迟蘖肥施用叶龄期(叶龄指数60%左右)、提前穗肥施用叶龄期(叶龄指数80%左右)同时增加穗肥施用比例,既可以显著提高氮素积累量、氮素吸收利用率、农学利用率及偏生产力,又能显著促进成穗率的提高和颖花数的分化,达到保蘖促花的双重作用,实现优源、扩库、充实的目标,从而获得高产。     

High night-time temperature during flowering and pod filling affects flower opening,yield and seed fatty acid composition in canola

Winter canola (Brassica napus L.) is highly sensitive to increasing temperatures during the reproductive and pod-filling stages. Although the impact of high day-time temperature stress on yield and quality has been documented in canola, similar information under high night-time temperature (HNT) stress is not available. Using six hybrids and four open-pollinated cultivars, we observed a marked shift in peak flowering towards earlier, cooler hours of the morning under HNT. Averaged across two independent experiments, the photochemical efficiency of photosystem II was significantly decreased (3%), with a significant increase in thylakoid membrane damage (13%) in the leaves of susceptible cultivars under HNT stress. Similarly, the susceptible cultivars also recorded significant reduction in biomass (34%), pod number (22%), pod weight (37%) and total seed weight (40%) per plant while the same set of agronomic traits were not affected among the tolerant cultivars. Quantitative impact of heat stress was confirmed with increased sensitivity to HNT exposure from gametogenesis until maturity resulting in a significantly higher yield loss compared to stress exposure from post-flowering till maturity. HNT significantly decreased oil concentration, but increased protein concentration and saturated fatty acid levels in seeds of the susceptible cultivars. However, HNT had no impact on the unsaturated fatty acids in both hybrids and the open-pollinated cultivars. Breeding targets based on fatty acid composition for enhancing canola seed quality may not be easily amenable due to the inconsistency documented with the compositional changes under heat stress. In summary, our findings conclude that canola hybrids are better suited to regions experiencing heat stress, compared to open-pollinated cultivars, indicating the possibility of a complete shift to hybrid canola cultivation under predicted hotter climates in the future.