Genes for resistance to powdery mildew in European barley cultivars registered in the Czech Republic from 2011 to 2015

Barley (Hordeum vulgare) is cultivated on 49.1 million hectares worldwide with 50.2% of the area located in Europe. Powdery mildew, caused by Blumeria graminis f. sp. hordei (Bgh), occurs wherever barley is grown. Cultivar resistance plays an important role in global barley production, especially in parts of Europe where high concentrations of both spring and winter types are grown. The aim of this report was to postulate specific resistance genes in barleys from nine European countries registered in the Czech Republic from 2011 to 2015. Thirty‐five spring cultivars and 27 winter barleys were tested with 56 diverse Bgh isolates. Twenty‐five known resistance genes were postulated, and unknown genes were detected in Sandra, Saturn and Zeppelin. Unidentified specific resistance genes were also present in winter hybrids Hobbit and Wootan. Spring cultivars Arthur and Francin consisted of three and two genotypes, respectively. Resistance gene mlo was present in 26 spring cultivars, and the proportion of cultivars with this gene increased from 62.9% in 2006–2010 to 75.7% in 2011–2015. The gene Mlp1 was identified for the first time in German winter cultivar Saturn. Five spring cultivars registered in Slovakia were included in the tests. All the cultivars that were tested contained one or more specific resistance genes to powdery mildew. Adaptability of the pathogen and possibilities for breeding winter barleys are discussed.     

寄主诱导的基因沉默提高植物真菌病害抗性研究进展

寄主诱导的基因沉默(host-induced gene silencing,HIGS)以病原菌生长发育、产孢繁殖、侵染或致病过程中的关键基因作为靶点,在寄主植物中表达针对靶基因的RNAi构建体,在病原菌侵染植物的过程中,摄入相应的ds RNA或si RNA分子,通过识别、结合特异核苷酸序列,干扰病菌靶基因表达,从而抑制病菌侵染和扩展,使植物表现抗病。这项技术为培育基于病原菌特异序列的植物抗病性奠定了基础,显示了巨大的应用潜力。本文综述了利用HIGS技术提高植物真菌病抗性的最新研究进展,总结了国内外利用这项技术改良植物真菌病害抗性的主要策略、技术路线,展望了发展应用前景。     

无核白葡萄多酚氧化酶基因的克隆与序列分析

为获得无核白葡萄中多酚氧化酶(PPO)的基因序列,以无核白葡萄果实为试材,进行基因同源克隆,得到多酚氧化酶基因CDS全长序列,其完整的编码框为1 824 bp,编码607个氨基酸(GenBank登陆号为KP271928);系统进化分析可知,其与葡萄科植物PPO基因同源性最高;生物信息学方法发现该蛋白分子量为67 392.44 Da,等电点为6.42,具典型的酪氨酸家族结构域,无信号肽结构,但有一个跨膜结构域,为亲水性蛋白;二级结构预测发现,PPO具有CuA与CuB两个结合区,分别嵌入蛋白活性腔中。该研究为进一步从分子水平探讨多酚氧化酶与褐变的关系奠定理论基础。     

东北黑土nirS型反硝化细菌群落和网络结构对长期施用化肥的响应

【目的】探明长期施用氮磷钾化肥对东北黑土农田土壤nirS型反硝化细菌群落和网络结构的影响,为更加合理的肥料配施提供理论依据。【方法】基于农业农村部黑龙江耕地保育与农业环境科学观测实验站平台,选取不施肥(NoF)、氮肥(N)、磷肥(P)、钾肥(K)、氮钾肥(NK)、氮磷肥(NP)、磷钾肥(PK)、氮磷钾肥(NPK)8个施肥处理,借助荧光定量PCR、Illumina MiSeq高通量测序和分子生态网络技术,分析东北黑土nirS型反硝化细菌丰度、群落及网络结构,及影响反硝化细菌群落变异的主要环境因子。【结果】1)长期施用氮肥均在显著增加nirS基因拷贝数的同时,降低了nirS型反硝化细菌的群落多样性,而磷、钾肥对其影响并不显著。2)Proteobacteria是所有处理中的优势反硝化细菌门,相对丰度为16.96%~27.34%,且氮肥的施用促进了隶属于该菌门中Bradyrhizobium的生长。3)PCoA分析结果显示,8个施肥处理nirS型反硝化细菌群落主要分成施氮肥和不施氮肥两组,说明长期氮肥的施用显著改变了东北黑土反硝化细菌的群落结构。结合Mantel test的结果可知,土壤pH是影响nirS型反硝化细菌群落改变的主要因素。4)分别构建施氮肥和不施氮肥的反硝化细菌分子生态网络,发现施氮肥和不施氮肥网络结构存在很大差异,长期施用氮肥明显简化了nirS型反硝化细菌的网络结构,同时使其网络结构稳定性降低,易受外界环境扰动。【结论】尽管施用化学氮肥有利于土壤养分的增加,但其土壤nirS型反硝化细菌群落及网络结构发生了较大改变,而磷、钾肥的施用对反硝化细菌群落无显著影响。本试验结果为进一步研究东北黑土区农田土壤反硝化微生物对不同施肥管理的响应机制提供了重要科学依据。     

施氮量和采收时期对油蔬两用型油菜菜薹维生素合成的影响

为明确施氮量和采收时期对油蔬两用型油菜薹维生素含量的影响,以富硒1号为材料,采用裂区试验设计,设置3个施氮量(120、180和240 kg·hm^-2)和4个采收时期(以株高20、30、40和50 cm表示),分析了油菜薹维生素C、维生素E、维生素B₁和维生素B₆含量的变化;比较了半乳糖内酯脱氢酶(GalLDH)、抗坏血酸氧化酶(AAO)、抗坏血酸过氧化物酶(APX)、脱氢抗坏血酸还原酶(DHAR)和单脱氢抗坏血酸还原酶(MDHAR)活性,并分析了维生素家族基因在菜薹中的表达量。结果表明,随着施氮量的增加,油菜薹还原型维生素C(AsA)含量降低;氧化型维生素C(DHA)含量在株高20 cm采收时增加,但在30 cm及以上株高采收时则降低;维生素E和维生素B₆含量先增加后降低;维生素B₁含量增加;维生素C合成关键酶GalLDH活性先略增加后降低;AAO和DHAR活性增加,APX和MDHAR活性则整体表现为先增加后降低。且施氮量增加后,维生素家族各基因在株高30和40 cm采收的菜薹中主要表现为正向调控,而在株高20和50 cm采收的菜薹中主要表现为负向调控。就采收时期而言,菜薹中的维生素含量在株高30~40 cm时高于其他时期;在同一施氮量下,随着菜薹采收株高的增加,维生素家族基因的表达量主要呈上调模式。综合分析可知,GalLDH活性的降低以及AAO/DHAR和APX/MDHAR代谢平衡是增施氮肥导致菜薹维生素C含量降低的重要原因。本研究结果为油蔬两用型油菜薹优质栽培技术提供了理论依据。     

Toxicity of cadmium and its competition with mineral nutrients for uptake by plants: A review

Cadmium(Cd)is a toxic heavy metal occurring in the environment naturally and is also generated through various anthropogenic sources and acts as a pollutant.Human health is affected by Cd pollution in farmland soils because food is the main source of Cd intake in the non-smoking population.For crops,Cd toxicity may result from a disturbance in uptake and translocation of mineral nutrients and disturbance in plant metabolism,inhibiting plant growth and development.However,plants have Cd tolerance mechanisms,including restricted Cd uptake,decreased Cd root-to-shoot translocation,enhanced antioxidant enzyme activities,and increased production of phytochelatins.Furthermore,optimal supply of mineral nutrients is one of the strategies to alleviate the damaging effects of Cd on plants and to avoid its entry into the food chain.The emerging molecular knowledge contributes to understanding Cd uptake,translocation,and remobilization in plants.In this review,Cd toxicity and tolerance mechanisms,agricultural practices to minimize Cd accumulation,Cd competition with essential elements(calcium,copper,iron,zinc,and manganese),and genes associated with Cd uptake are discussed in detail,especially regarding how these mineral nutrients and genes play a role in decreasing Cd uptake and accumulation in crop plants.     

茶树CsSNAT基因的克隆与表达分析

为明确茶树SNAT基因的序列特征和表达特点,在茶树转录组测序的基础上以茶树(Camellia sinensis)品种舒茶早为试验材料,通过SMART^TM RACE技术克隆茶树褪黑素合成途径中关键限速酶5-羟色胺-N-乙酰转移酶CsSNAT基因的cDNA全长序列,并分析其基因结构和表达模式。结果表明,CsSNAT基因序列全长1 014 bp,其中包含742 bp的完整开放阅读框(ORF),编码247个氨基酸,预测等电点7.64,蛋白分子量27.36 kDa。氨基酸序列比对显示,CsSNAT蛋白与其他高等植物的SNAT蛋白具有较高同源性,与葡萄 VvSNAT(CBI31163)同源性最高,为66.19%。诱导蛋白最佳条件为温度28℃,异丙基-β-D-硫代半乳糖苷(IPTG)浓度0.5 mg·L^-1,诱导时间6 h,在上清液中可获得大量分子量为66 kDa的可溶性融合蛋白。实时定量 PCR 分析表明,CsSNAT在褪黑素处理6 h后表达量最高,为对照的3倍;茶尺蠖取食6 h后,CsSNAT表达显著上调。不同激素处理结果显示,CsSNAT受脱落酸(ABA)、茉莉酸甲酯(MeJA)和水杨酸(SA)诱导表达。本研究结果为5-羟色胺-N-乙酰转移酶功能研究奠定了一定的基础。     

谷子MDH基因非生物逆境响应特性研究

为揭示谷子MDH基因对逆境胁迫的响应,本研究通过序列比对得到2个谷子苹果酸脱氢酶(malate dehydrogenase, MDH)基因SiMDH1、SiMDH2,并对SiMDH1和SiMDH2的编码蛋白特征、基因结构、功能、进化等性状进行了系统的分析和预测,用荧光实时定量PCR(qPCR)检测了SiMDH1和SiMDH2在苗期不同逆境及关键生育期干旱胁迫和不同光照强度下的表达。蛋白特征参数和序列分析表明,SiMDH1和SiMDH2蛋白特征参数相似度很高,序列非常保守,都含有MDH基因的特征功能域苹果酸酶NAD结合域和苹果酸酶C端功能域。亚细胞定位预测显示,SiMDH1和SiMDH2主要定位于叶绿体、细胞质和线粒体。启动子区域顺式元件分析发现,在SiMDH1和SiMDH2启动子区域含有大量光应答、激素类和其他类应答元件。苗期逆境qPCR表达谱分析发现,SiMDH1和SiMDH2受脱落酸(ABA)、聚乙二醇(PEG)、高盐和低温不同程度诱导,揭示SiMDH1和SiMDH2参与了谷子苗期非生物逆境胁迫应答。进一步分析发现,SiMDH1和SiMDH2参与了谷子在拔节、抽穗、灌浆期的干旱应答,而光照强度会显著影响SiMDH1和SiMDH2基因在不同生育期的表达。本研究为进一步分析MDH逆境应答机制以及利用基因工程方法改善作物抗逆性提供了试验和理论支持。     

Dynamics of nitrogen translocation from mature leaves to new shoots and related gene expression during spring shoots development in tea plants (Camellia sinensis L.)

The contribution of N remobilization is crucial for new shoots growth and quality formation during spring tea shoots development. However, the translocation mechanism of N from source leaves to sink young shoots is not well understood. In the present study, ¹⁵N urea was applied to mature tea leaves one week before bud break to track N remobilization in a field experiment. The dynamic changes in plant ¹⁵N abundance, contents of amino acids, and the expression levels of genes related to N metabolism and translocation were followed during the 18‐d development of new spring shoots until three expanding young leaves. The results showed that during the growth of new shoots the amount of ¹⁵N in the shoots increased, whereas the N_dff (N derived from ¹⁵N‐urea) in mature leaves decreased, showing that the foliar‐applied N in mature leaves was readily exported to new shoots. This process was found to be accompanied by decline of chlorophylls. In the mature leaves, expression CsATG18a and CsSAG12 involved in autophagy was dramatically induced (> 4‐fold) at approximately nine days after the bud breaking. The genes involved in the transformation of amino acids, including primarily CsGDH2, CsGDH4, CsGLT3, CsGS1;3, and CsASN2 were upregulated by > 3‐fold after bud breaking. The expression levels of CsATG8A, CsATG9, CsSAG12, CsGS1;1, CsGDH1, and CsAAP6 correlated negatively with the N_dff in mature leaves, but positively with ¹⁵N amount and total N amount in new shoots, suggesting these genes played important roles in N export from mature leaves. In the new shoots, the expression of most genes showed two defined peaks, one on six days and one on 12 days after bud breaking. The expression of CsGS2, CsASN3, CsGLT1, and CsAAP4 positively correlated with the ¹⁵N amount and total N amount in new shoots. These genes might be involved in the transport and re‐assimilation of N from mature leaves. The overall results demonstrated that the translocation of ¹⁵N from mature leaves to new spring shoots was regulated by the genes involved in autophagy, protein degradation, amino acid transformation and transport.