椰子组培研究四十年

组织培养对于椰子快速繁殖以及全球椰子产业发展具有重要意义。本文就40年来国内外在椰子组培快繁技术,包括椰子组培技术的重要性,不同外植体如胚芽、花序、子房等在组织培养中的应用与优劣,不同培养基和培育环境对组培效果的影响等要素,进行了详细的总结和讨论。同时总结了椰子组培快繁技术的进展,分析了目前限制椰子组培快繁技术发展的因素,提出了应对策略与具体建议。本文对椰子快繁技术及椰子种质资源保存具有参考价值。     

外源ABA对苹果砧木叶片解剖结构 及内源激素含量的影响

通过光镜和透射电镜分别观察外源脱落酸（ABA）处理下楸子（Malus prunifolia）、平邑甜茶（M. hupehensis）和新疆野苹果（M. sieversii）叶片解剖结构及叶绿体超微结构的变化，利用扫描电镜和酶联免疫法分别研究ABA处理对叶片气孔特征及其内源激素含量的影响。结果表明：与对照相比，ABA处理下楸子、平邑甜茶和新疆野苹果的叶厚分别减少了7.93%、0.25%和0.81%，栅栏组织厚度分别减少了31.43%、8.53%和4.99%（P＜0.05），海绵组织厚度分别增加了10.34%、6.14%和5.63%（P＜0.05），叶肉组织结构疏松度（SR）分别增加了19.59%、6.55%和6.50%。ABA处理下楸子和平邑甜茶的栅栏组织/海绵组织厚度比值（P/S）及叶肉组织结构紧密度（CTR）较对照显著减少（P＜0.05），其中，P/S值分别下降37.86%和13.82%，CTR分别下降25.46%和8.29%，而新疆野苹果的P/S值和CTR下降但不显著。此外，ABA处理下楸子和平邑甜茶的上表皮细胞厚度较对照分别增加了5.82%和6.43%，新疆野苹果的较对照减少了26.23%（P＜0.05）；楸子和新疆野苹果的下表皮细胞厚度较对照增加了12.09%和14.21%（P＜0.05），平邑甜茶的较对照减少了12.56%。平邑甜茶和新疆野苹果叶片上下角质层厚度较对照显著增加（P＜0.05），而楸子的变化不显著。在ABA诱导下，3种砧木的气孔密度、气孔大小（长度×宽度）及其开口度和开张比均不同程度地下降，其中，楸子的较对照分别下降了3.62%、7.12%×19.59%、67.60%和86.66%，平邑甜茶的分别下降了3.50%、4.99%×20.65%、32.42%和58.24%，新疆野苹果的分别下降了8.54%、0.92%×12.06%、20.37%和16.35%。ABA处理下3种砧木叶片细胞中叶绿体的数量变少，类囊体结构排列疏松，叶绿体上的淀粉粒趋于变小。外施ABA使3种砧木叶片内源ABA和玉米素核苷（ZR）的含量极显著增加（P＜0.01），其中楸子ABA和ZR含量比对照分别增加30.83%和13.31%，平邑甜茶的分别增加62.40%和45.28%，而新疆野苹果的分别增加了37.07%和17.06%。楸子和新疆野苹果叶片的吲哚乙酸（IAA）和赤霉素（GA）含量无显著变化，而平邑甜茶叶片的IAA及GA含量比对照分别增加了62.62%和20.62%（P＜0.01）。总之，在ABA处理下，3种苹果砧木的叶片组织解剖结构和气孔特征都发生旱生性结构变化，叶肉细胞中淀粉粒趋于变小，叶片内源ABA和ZR水平增加显著，而IAA和GA水平变化因基因型不同而存在差异。     

高粱光敏色素A基因的克隆、蛋白结构与长短日照诱导表达模式

光敏色素是一类红光/远红光受体,对植物光形态建成、外部形态和生理功能等方面起着重要的调节作用。为明晰光敏色素A在调控高粱光形态建成发育过程中的作用,本研究比较分析光温敏感程度不同的高粱品种Btx623、Rio、Tx430和LH645的PhyA基因序列和PHYA蛋白的保守结构域,利用qRT-PCR明确长短日照条件处理下PhyA基因的表达模式。结果表明,Btx623、Rio和Tx430之间PhyA基因序列存在非同义突变,LH645在PhyA基因的第7个外显子上插入了5 bp碱基序列;高粱PhyA蛋白中含有1个PAS-2结构域、1个GAF结构域、1个Phytochrome结构域、2个PAS结构域、1个组氨酸激酶A结构域和1个类似组氨酸激酶的ATP激酶结构;遗传进化树分析表明,单子叶和双子叶植物PhyA蛋白明显聚为2大类,高粱PhyA、谷子PhyA和玉米PhyA1、PhyA2聚为一个亚类,相互间亲缘关系较近,与水稻PhyA同源性相对较远;在短日照(SD)条件下各时期PhyA基因转录水平变化显著,Rio PhyA基因转录水平较Btx623和Tx430变化明显,LH645 PhyA基因转录变化不显著,在长日照(LD)条件下Rio PhyA基因转录水平低于Btx623和Tx430,LH645 PhyA基因转录水平变化不显著。以上结果表明,PhyA受光周期诱导,推测其在高粱光形态建成中起重要调控作用。本研究为进一步解析PhyA基因功能及其在调控高粱光形态建成发育过程中的作用提供理论基础。     

蛋鸡育成期和产蛋高峰期生殖激素水平及激素受体基因表达规律研究

【目的】探究蛋鸡育成期和产蛋高峰期相关生殖激素含量及其受体基因mRNA表达量在24 h内的变化规律与差异,分析血液生殖激素含量变化与产蛋排卵行为的关系。【方法】以育成期和产蛋高峰期的罗曼蛋鸡为研究对象,监测其在产蛋高峰期的产蛋时间点,运用酶联免疫吸附法(ELISA)测定2个时期蛋鸡血液中前列腺素(PGs)、雌二醇(E2)、促卵泡素(FSH)、黄体生成素(LH)含量在24 h内的动态变化,实时荧光定量PCR技术检测相应时间点的前列腺素F受体(PTGFR)、雌激素受体(ERα)、促卵泡素受体(FSHR)和黄体生成素受体(LHR)基因mRNA在输卵管子宫部的日表达水平。【结果】蛋鸡产蛋时间集中在早上08:00-10:00。产蛋高峰期时,一天中06:00、10:00、14:00、18:00的PGs含量显著高于育成期(P0.05),E2含量在06:00和18:00极显著高于育成期(P0.01),FSH含量在18:00显著高于育成期(P0.05),LH含量在14:00、22:00和02:00显著高于育成期(P0.05)。育成期除LH外,其余3种激素含量在一天中波动不大。产蛋高峰期PGs含量在产蛋前显著高于产蛋后,PTGFR mRNA表达量在产蛋前后4 h均显著(P0.05)升高;FSH和E2含量在产蛋后(18:00)极显著(P0.01)上升。子宫组织的激素受体基因表达量变化与相应血液激素含量变化一致。【结论】蛋鸡产蛋时间集中于上午,产蛋高峰期各生殖激素含量总体高于育成期,蛋鸡育成期和产蛋高峰期的血液生殖激素水平变化规律与输卵管子宫部激素受体基因mRNA表达量的变化规律基本一致。PGs参与蛋鸡产蛋高峰期的产蛋和排卵过程,E2和FSH可能在蛋壳形成过程中发挥调控作用。     

罹患口腔炎乌苏里蝮的相关生理学指标分析

口腔炎是人工养殖蛇类的常见病之一,为了深入了解和尽早做好预防工作,对罹患口腔炎的成年雄蛇乌苏里蝮(Gloydius ussuriensis)进行解剖观察,并检测了相关器官的系数、血液学指标和激素等。结果表明:整体肥满度、躯体肥满度、肾系数等数值罹患口腔炎蛇略高于健康蛇;而肝系数、精巢系数和脂肪体系数等罹患口腔炎的蛇低于健康雄蛇,且前两者呈显著性差异,后者呈极显著性差异。性激素检测结果显示罹患口腔炎的蛇血液睾酮和雌二醇含量均低于健康蛇,且均具极显著性差异。血细胞指标中,红细胞数量、红细胞分布均差、白细胞总数、淋巴细胞数量和淋巴细胞比率等均为罹患口腔炎的蛇高于健康雄蛇,且具显著性差异,但嗜中性粒细胞和淋巴细胞的比值(NLR)、中性粒细胞数量和比率、中间细胞数量及其比率均为患口腔炎的雄蛇低于健康蛇,且具显著性差异。结论:口腔炎不仅影响了主要代谢器官的器官系数的变化,而且血液多项指标和性激素水平也有明显差异。性激素含量的变化将对乌苏里蝮的正常生殖繁育产生不可忽视的影响。另外,NLR和红细胞分布宽度2个血常规指标可作为早期诊断炎症发生的首选参数。     

Sewage sludge processing determines its impact on soil microbial community structure and function

Composting and thermal drying are amongst the most commonly used post-digestion processes for allowing sanitation and biological stabilization of sewage sludge from municipal treatment plants, and making it suitable as soil conditioner for use in agriculture. To assess the impact of sludge-derived materials on soil microbial properties, fresh (LAF), composted (LAC) and thermally dried (LAT) sludge fractions, each resulting from a different post-treatment process of a same aerobically digested sewage sludge, were added at 1% (w/w) application rate on two contrasting (a loam and a loamy sand) soils and incubated under laboratory conditions for 28 days. Soil respiration, microbial ATP content, hydrolytic activities and arginine ammonification rate were monitored throughout the incubation period. Results showed that soil biochemical variables, including the metabolic quotient (qCO₂), were markedly stimulated after sludge application, and the magnitude of this stimulatory effect was dependent on sludge type (precisely LAT > LAF > LAC), but not on soil type. This effect was related to the content of stable organic matter, which was lower in LAT. Genetic fingerprinting by PCR–DGGE revealed that compositional shifts of soil bacterial and, at greater extent, actinobacterial communities were responsive to the amendment with a differing sludge fraction. The observed time-dependent changes in the DGGE profiles of amended soils reflected the microbial turnover dependent on the sludge nutrient input, whereas no indications of adverse effects of sludge-borne contaminants were noted. Our findings indicate that composting rather thermal drying can represent a more appropriate post-digestion process to make sewage sludge suitable for use as soil conditioner in agriculture.     

肉苁蓉提取物对雄性小鼠性激素分泌的影响

肉苁蓉化学成分复杂,有多种活性作用。本试验利用超声波协助提取、酸提取、醇提取方法提取肉苁蓉生物碱,分别对小鼠进行灌胃给药,观察肉苁蓉提取物对小鼠性激素分泌的影响。结果显示,乙醇提取方法得率最高,肉苁蓉提取物能显著提高小鼠雄激素含量（P〈0.05）。     

激素对奶牛乳腺上皮细胞乳脂肪合成的调控作用

乳脂肪是高质量的天然脂肪，其可为人类提供营养和能量，在各种膳食脂肪和油类中，是最容易被消化吸收的。乳脂肪是在乳腺中由从头合成或外源摄取的脂肪酸与甘油酯化形成的一种脂类物质，其含量的高低关系着牛奶品质的优劣和乳制品的加工特性。在奶牛的泌乳周期中，乳腺泌乳功能受多种因素影响，其中内分泌腺分泌的多种激素对奶牛乳腺上皮细胞（BMECs）乳脂的合成具有积极的调控作用。综上所述，作者介绍了氢化可的松、催乳素、胰岛素和生长激素4种泌乳相关激素对BMECs乳脂肪合成的调控机理，即从乳脂合成适宜的激素添加量、激素对乳脂球形态的影响方面初步阐释其调控作用，并从乳脂合成的关键酶及转录因子、激素对乳脂合成相关基因表达量方面深入阐释其作用机理，旨在为研究泌乳相关激素对奶牛乳腺内乳脂肪合成的调控机理提供参考。     

Gene cloning of ZmMYB59 transcription factor in maize and its expression during seed germination in response to deep‐sowing and exogenous hormones

Discovering the key genes responsible for deep‐sowing germination is very meaningful to cultivate maize varieties with strong germination ability in soil. In this study, ZmMYB59 gene whose functions remain unknown was successfully cloned. Subcellular localization showed that ZmMYB59 was localized in the nucleus. By analysing cis‐acting elements of its promoter sequence, many elements of MYB binding domain responsive to hormones (particularly gibberellin, GA) were found. On this basis, ZmMYB59 expression in different sowing depths, germination stages, tissues and treatments was analysed by real‐time PCR. ZmMYB59 expression levels in embryos or seedlings were significantly decreased with the increase in sowing depth at 2 days or 4 days after sowing (DAS). Further analysis showed that its expression in mesocotyls was also significantly downregulated with the increase in sowing depth at both 6 DAS and 8 DAS. Similarly, seed soaking with 10^‐5 M GA₃ inhibited ZmMYB59 expression in mesocotyls at 3 DAS. These results suggest that ZmMYB59 may play a negative regulatory role in seed germination in deep soil and the regulation is involved in GA signalling pathway.     

Muscular hypertrophy and growth‐promoting effects in juvenile pejerrey (Odontesthes bonariensis) after oral administration of recombinant homologous growth hormone obtained by a highly efficient refolding process

Growth hormone (GH) can be orally administrated to fish in order to increase growth rates. Fish growth is characterized by the hyperplasia and hypertrophy of muscle fibre throughout adult life. In this respect, GH could affect directly and indirectly (by growth and metabolic factors) the development and growth of muscle fibres. Recombinant pejerrey GH (r‐pjGH) was expressed in Escherichia coli and refolded in a highly efficient batch dilution system, obtaining 0.1 g L^?1 of hormone without protein precipitation during the refolding procedure. Orally administered hormone to pejerrey produced a 30% increase in mean weight and stimulated liver insulin‐like growth factor type I (IGF‐I) mRNA expression after 1 month of treatment. Histological analyses showed that muscle growth was generated mainly by hypertrophy of the fibres. A higher r‐pjGH dose increased muscle fibre hypertrophy but somatic growth was negatively affected probably due to a reduced capacity of generating new fibres.