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.     

增施有机肥改善黑土物理特性与促进玉米根系生长的效果

为改善东北黑土区粘重耕层的土壤物理特性,通过玉米秸秆还田基础上增施有机肥试验,拟明确增施有机肥对黑土物理特性和根系生长的提升效果。利用2015～2018年吉林省公主岭市和黑龙江省克山县黑土区的定位试验,测定了玉米抽雄期3种秸秆还田处理及其增施有机肥(旋耕秸秆还田+有机肥RSM、深翻秸秆还田+有机肥DSM、深松秸秆还田+有机肥SSM)处理的土壤物理指标;并采用微根管法原位测定了根系生长指标,计算出增施有机肥后各土壤物理特性与根系生长指标的变化量。结果表明,相比秸秆还田处理,增施有机肥降低了土壤容重、土壤紧实度,提升了土壤含水量,同时根长密度、根尖数密度和根平均直径均显著增加,其中根长密度和根尖数密度各土层平均增加了0.18 cm/cm~2和34.9×10~(-3)个/cm~2。不同秸秆还田方式增施有机肥后对黑土物理特性和根系生长的改善效果不同,其中0～15 cm土层RSM处理改善效果最明显,15～45 cm土层SSM和DSM处理改善效果最明显。有机肥和秸秆还田方式互作对黑土物理特性和促进根系生长指标具有显著的正向互作效应。上述结果表明,深松秸秆还田和深翻秸秆还田基础上增施有机肥模式更有利于改善黑土物理特性和促进根系生长,是改善东北黑土区粘重耕层的技术选择。     

不同温度下TMV侵染枯斑三生烟的LncRNA差异表达

【目的】 筛选不同温度下烟草花叶病毒（Tobacco mosaic virus,TMV）侵染后枯斑三生烟（Nicotiana tabacum var. Samsun NN）差异表达的长链非编码RNA（long non-coding RNA,lncRNA）,研究lncRNA在枯斑三生烟抗性反应中的作用。【方法】 N基因的温度敏感性使枯斑三生烟在25℃时具备对TMV的抗性、在31℃抗性丧失,在这两个温度条件下对枯斑三生烟接种TMV和磷酸盐缓冲盐水（phosphate buffered saline,PBS）,48 h后提取系统叶总RNA,构建链特异性文库后进行深度测序。对测序结果进行过滤后利用HTSeq将有效数据与近缘品种TN90（N. tabacum var. TN90）基因组比对,筛选得到lncRNA后利用FPKM法估计lncRNA的表达水平。通过edgeR筛选差异表达lncRNA（differentially expressed lncRNA,DElncRNA）,并利用qRT-PCR技术对这一结果进行验证。通过共定位及共表达分析预测DElncRNA的靶基因,通过参考基因组注释、GO和KEGG富集分析研究靶基因的功能。【结果】 4个处理共12个样本经lncRNA-seq各测得约8 000万条clean reads,共获得4 737条已知lncRNA、40 169条新lncRNA。其中64个lncRNA在不同温度条件下TMV侵染后存在差异表达,qRT-PCR测定结果显示这些lncRNA的测序正确率在80%左右,表明本研究所得测序数据具备较高的可信度。对DElncRNA进行靶基因预测,发现一些基因同时被25℃下调和31℃上调的DElncRNA靶向。靶基因注释功能丰富,主要参与植物抗病、激素和代谢等生理过程。部分可能与激素通路相关的lncRNA,在25℃下TMV侵染时呈现下调趋势,而在31℃下TMV侵染则呈现上调趋势。GO富集分析显示靶基因主要参与构成膜、囊泡等组分,具备钙、钾离子通道抑制剂活性等分子功能,使相应离子得以转运引发随后的反应,同时也参与发病、抗原加工和呈现、细胞分裂素代谢等生理过程。KEGG分析发现靶基因显著富集在植物激素信号转导通路,25℃下调和31℃上调的DElncRNA靶基因同时富集在激素信号传导、ABC运输蛋白、苯丙烷类生物合成等通路。【结论】 不同温度（25℃和31℃）条件下TMV侵染枯斑三生烟后,长链非编码RNA差异表达,DElncRNA通过作用于激素信号传导、物质转运等过程参与寄主系统获得性抗性反应。研究结果可为揭示植物系统获得性抗性中lncRNA的调控功能以及新型抗病毒技术开发提供依据。     

甘蓝型油菜对华北坝上冷凉环境的适应性

针对华北坝上冷凉区短季类经济作物缺乏,芥菜型油菜品质差、产量低等问题,于2017-2018年在农业农村部张北农业资源与生态环境重点野外观测试验站,引种了甘蓝型双低油菜品种华油杂62,以当地传统芥菜型油菜品种大黄为对照,监测了两品种的叶面积指数、角果净光合速率日变化、地上部干物质积累量、养分积累量和油菜农艺性状、产量等。结果表明,华油杂62较大黄生育期长约10d,平均叶面积指数高23.36%~36.73%;角果净光合速率不及大黄,N、P、K积累量高于大黄;干物质快速增长期延长21d,干物质积累潜值高46.86%;菜子产量较大黄高22.8%~54.0%。甘蓝型油菜华油杂62对华北坝上冷凉区的地理气候环境有较好的生态适应性,具有替代区域传统芥菜型油菜的优势。     

Screening of antagonistic Trichoderma strains and their application for controlling stalk rot in maize

Maize is one of the major crops in China, but maize stalk rot occurs nationwide and has become one of the major challenges in maize production in China. In order to find an environment-friendly and feasible technology to control this disease, a Trichoderma-based biocontrol agent was selected. Forty-eight strains with various inhibition activities to Fusarium graminearum, and Fusarium verticillioides were tested. A group of Trichoderma strains(DLY31, SG3403, DLY1303 and GDFS1009) were found to provide an inhibition rate to pathogen growth in vitro of over 70%. These strains also prevented pathogen infection over 65% and promoted the maize seedling growth for the main root in vivo by over 50%. Due to its advantage in antifungal activity against pathogens and promotion activity to maize, Trichoderma asperellum GDSF1009 was selected as the most promising strain of the biocontrol agent in the Trichoderma spectrum. Pot experiments showed that the Trichoderma agent at 2–3 g/pot could achieve the best control of seedling stalk rot and promotion of maize seedling growth. In the field experiments, 8–10 g/hole was able to achieve over 65% control to stalk rot, and yield increased by 2–11%. In the case of natural morbidity, the control efficiency ranged from 27.23 to 48.84%, and the rate of yield increase reached 11.70%, with a dosage of Trichoderma granules at 75 kg ha~(-1). Based on these results, we concluded that the Trichoderma agent is a promising biocontrol approach to stalk rot in maize.     

亚洲玉米螟幼虫膜结合海藻糖酶基因RNAi效应

海藻糖是昆虫的血糖，为昆虫提供能量；海藻糖酶催化海藻糖分解为葡萄糖，是几丁质生物合成的原料。围食膜（peritrophic membranes，PMs）是昆虫消化道特有结构，对昆虫消化食物、保护肠道表皮细胞具有重要作用；几丁质是PMs的重要组分。海藻糖酶（trehalase，Tre）和几丁质合成酶（chitin synthase，CHS）是几丁质合成途径的第1个和最后1个酶。本研究通过饲喂亚洲玉米螟（Orstrinia furnacalis，Asian corn borer，ACB）膜结合海藻糖酶（OfMT）基因特异的干扰dsRNA，研究RNAi对ACB幼虫中肠CHSB（OfCHSB）基因表达及幼虫发育的影响。发现处理48 h后，OfMT基因和OfCHSB基因表达量分别下降了52%和53%，幼虫发育迟缓。通过对处理及对照ACB幼虫中肠石蜡切片进行苏木精-伊红染色（hematoxylin-ethanol，HE染色）和几丁质标记，发现血腔内脂肪体组织减小、中肠围食膜组织中几丁质含量减少。推测OfMT基因有可能成为ACB的生物防治的靶标基因。     

Heterologous expression of the ThIPK2 gene enhances drought resistance of common wheat

Th IPK2 is an inositol polyphosphate kinase gene cloned from Thellungiella halophila that participates in diverse cellular processes. Drought is a major limiting factor in wheat(Triticum aestivum L.) production. The present study investigated whether the application of the Th IPK2 gene could increase the drought resistance of transgenic wheat. The codon-optimized Th IPK2 gene was transferred into common wheat through Agrobacterium-mediated transformation driven by either a constitutive maize ubiquitin promoter or a stress-inducible rd29 A promoter from Arabidopsis. Molecular characterization confirmed the presence of the foreign gene in the transformed plants. The transgenic expression of Th IPK2 in wheat led to significantly improve drought tolerance compared to that observed in control plants. Compared to the wild type(WT) plants, the transgenic plants showed higher seed germination rates, better developed root systems, a higher relative water content(RWC) and total soluble sugar content, and less cell membrane damage under drought stress conditions. The expression profiles showed different expression patterns with the use of different promoters. The codon-optimized Th IPK2 gene is a candidate gene to enhance wheat drought stress tolerance by genetic engineering.     

软基水闸底板脱空动力学反演模型构建与试验验证

软基水闸极易因地基不均匀沉降、渗透变形等发生底板脱空现象，传统的无损检测方法难以在水下探测底板脱空，且不具实时性，其应用还存在一定局限性。该研究基于软基水闸室内物理模型，提出了一种软基水闸底板脱空动力学反演方法。首先，基于多参数变量的底板脱空范围数学模型，改进了反映软基水闸底板脱空参数与水闸动力学参数（模态参数）之间非线性映射关系的数学代理模型；其次，以水闸多测点多阶频率和振型变化率组合作为动力学敏感特征量，建立了软基水闸底板脱空参数反演的目标函数，并基于遗传算法对目标函数进行求解。最后，建立了软基水闸室内物理模型，并在模型中设置3种不同的底板脱空工况，采用软基水闸底板脱空动力学反演方法对软基水闸底板的脱空进行反演识别与模型验证。结果表明：基于动力学参数反演识别的水闸底板脱空区域趋势和脱空面积与模型实际脱空情况吻合较好，3种工况下水闸底板脱空面积反演结果与模型实际脱空面积的相对误差分别为7.47%、6.78%、6.90%，验证了该方法的可靠性，可望为软基水闸实际工程底板脱空隐患检测提供一种新的思路。