甜瓜VQ家族基因鉴定及白粉病菌响应分析

VQ家族基因是植物特有的一类基因,其编码蛋白主要与WRKY蛋白相互作用协同调控下游基因表达,在植物生长发育及抵御各种外界环境胁迫中起重要作用。为揭示甜瓜VQ家族基因在抗白粉病中的功能,本研究采用生物信息学方法从甜瓜基因组中鉴定到26个VQ家族基因,这些基因不均匀地分布在甜瓜11条染色体上。系统进化分析表明,该家族成员分布于不同的分支中,分别与拟南芥VQ家族蛋白不同成员聚在一起。基因结构分析表明,甜瓜VQ家族基因序列均无内含子,只有1个外显子。组织表达分析表明,1个甜瓜VQ家族基因组成性表达,23个甜瓜VQ家族基因组织特异性表达。此外,白粉病菌侵染之后,6个VQ家族基因呈现不同程度的响应。本研究结果为进一步研究甜瓜VQ家族基因在甜瓜抗白粉病中的功能奠定了理论基础。     

Modelling soil erodibility in mountain rangelands of southern Kyrgyzstan

Soil erosion in mountain rangelands in Kyrgyzstan is an emerging problem due to vegetation loss caused by overgrazing. It is further exacerbated by mountain terrain and high precipitation values in Fergana range in the south of Kyrgyzstan. The main objective of this study was to map soil erodibility in the mountainous rangelands of Kyrgyzstan. The results of this effort are expected to contribute to the development of soil erodibility modelling approaches for mountainous areas. In this study, we mapped soil erodibility at two sites, both representing grazing rangelands in the mountains of Kyrgyzstan and having potentially different levels of grazing pressure. We collected a total of 232 soil samples evenly distributed in geographical space and feature space. Then we analyzed the samples in laboratory for grain size distribution and calculated soil erodibility values from these data using the Revised Universal Soil Loss Equation (RUSLE) K-factor formula. After that, we derived different terrain indices and ratios of frequency bands from ASTER GDEM and LANDSAT images to use as auxiliary data because they are among the main soil forming factors and widely used for prediction of various soil properties. Soil erodibility was significantly correlated with channel network base level (geographically extrapolated altitude of water channels), remotely sensed indices of short-wave infrared spectral bands, exposition, and slope degree. We applied multiple regression analysis to predict soil erodibility from spatially explicit terrain and remotely sensed indices. The final soil erodibility model was developed using the spatially explicit predictors and the regression equation and then improved by adding the residuals. The spatial resolution of the model was 30 m, and the estimated mean adjusted coefficient of determination was 0.47. The two sites indicated different estimated and predicted means of soil erodibility values (0.035 and 0.039) with a 0.05 significance level, which is attributed mainly to the considerable difference in elevation.     

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.     

长期秸秆还田和地下水位对土壤镉积累及有效性的影响

以1982年开始的长期定位试验为对象,研究了2个地下水位(高水位-20 cm和低水位-80 cm)下3种施肥方式(高量秸秆还田HS、常量秸秆还田MS、化肥CF)对水稻土镉积累及有效性的影响。结果表明:高水位条件下,长期秸秆还田可增加土壤总镉及有效态镉含量,HS和MS处理分别较CF处理提高21.8%、59.9%和9.8%、49.2%。低水位条件下,HS处理土壤总镉及有效态镉含量高于CF处理,较CF处理分别提高11.2%和31.6%; MS处理土壤总镉及有效态镉含量低于CF处理,较CF处理分别下降8.8%和14.3%。2012年,在保证原定位试验有足够重复的前提下,变更高水位条件下的部分试验处理。当HS处理变更为CF处理5 a后,土壤总镉和有效态镉含量分别较变更前下降2.5%和5.7%;CF处理变更为MS处理后,土壤总镉和有效态镉含量分别较变更前增加16.5%和58.9%。相同施肥处理,高水位土壤总镉含量高于低水位土壤。在MS处理下,高水位土壤有效态镉含量也高于低水位土壤,而在HS和CF处理下,低水位土壤有效态镉含量高于高水位土壤。土壤镉含量和土壤基本理化性质的相关分析表明,土壤总镉与土壤有机质、络合态铁含量极显著正相关,与pH显著负相关;土壤有效态镉与土壤pH、游离氧化铁极显著负相关。研究表明,施肥方式和地下水位是通过改变土壤的基本性质影响土壤镉的积累及其有效形态。     

芜菁Br14-3-3的克隆及其在非生物胁迫下的表达分析

克隆了‘津田芜菁’（Brassica rapa subsp. rapifera‘Tsuda’）通用调节因子14-3-3基因cDNA序列，命名为Br14-3-3（GenBank登录号为MK896872）。该基因全长1 113 bp，开放阅读框全长774 bp，编码含有257个氨基酸的多肽。荧光定量PCR分析Br14-3-3在‘津田芜菁’不同组织中及其在温度、脱水、渗透、ABA和无机盐等非生物胁迫下幼苗中的表达，结果表明该基因在‘津田芜菁’的花中表达量最高，幼苗中次之；低温抑制了Br14-3-3的表达，其他非生物胁迫可诱导该基因表达，暗示Br14-3-3在非生物胁迫应答中发挥功能。     

枸杞Lb_PPR1的克隆及其在不同育性品种中的表达分析

三角状五肽重复（Pentatricopeptide repeats，PPR）蛋白定位于多种细胞器中，参与细胞核和细胞器中特异单链RNA的转录后修饰和编辑，在植物生长发育的多个阶段均发挥着重要的作用。分别从枸杞（Lycium barbarum）雄性可育系‘宁杞1号’和不育系‘宁杞5号’中克隆了Lb_PPR1基因，并对其编码蛋白质进行理化性质、亚细胞定位、保守结构域、蛋白结构以及系统进化等方面进行预测分析。结果显示，‘宁杞1号’和‘宁杞5号’中Lb_PPR1基因的开放阅读框均为1 977 bp，编码658个氨基酸，但其中存在20个碱基和14个氨基酸的差异。Lb_PPR1蛋白包含14个串联重复的PPR保守基序，属于PLS家族，该蛋白定位在细胞膜和细胞质。二级、三级结构预测显示该蛋白以α螺旋为主。同源进化分析得出Lb_PPR1蛋白与同属茄科的辣椒和烟草PPR蛋白亲缘关系最近。利用实时荧光定量PCR检测Lb_PPR1在枸杞不同组织器官及不同发育阶段花药中的表达特性。结果显示，Lb_PPR1在枸杞不同组织中均有表达，但在可育系花蕾中表达量最高，不育系‘宁杞5号’各组织中表达量均显著低于可育系‘宁杞1号’。以上结果表明，Lb_PPR1基因可能与枸杞花药发育有关。     

花鲈cox6a基因与相关miRNA的鉴定及其在盐度适应中的表达调控分析

细胞色素c氧化酶（COX）是线粒体电子传递链的末端酶，COX6A是细胞色素c氧化酶的核编码亚基之一。为丰富花鲈（基因及相关miRNAs在花鲈渗透调节机制中的潜在作用，本研究开展了花鲈cox6a基因的miRNAs，并对其与）。系统进化树分析进一步确认了两个花鲈在垂体、脑、肝脏和肾脏中的表达量较高，基因的miRNA：miR-30e-5p、miR-223、miR-200a-3p和miR-155-5p，但没有发现可能靶定基因以及miR-30e-5p、miR-223、miR-200a-3p都呈现显著差异表达，而miR-155-5p仅在适应高盐环境时出现了显著的差异表达。这表明，基因和4个miRNAs在花鲈的渗透调节过程中发挥潜在作用。其中miR-30e-5p、miR-223和miR-200a-3p的表达趋势与cox6a2基因的表达调控。本研究为探讨花鲈适应不同盐度环境的渗透调节机制提供了基础数据。     

栉孔扇贝Dmrt1的分子鉴定及表达模式分析

DMRT1是DMRT家族重要成员之一，主要参与动物性别决定和分化调控，但在不同动物中其表达和功能调控作用存在差异。本研究采用生物信息学方法分析了栉孔扇贝（Chlamys farreri）Dmrt1的序列特征，采用半定量RT-PCR技术确定了其mRNA在成体性腺、闭壳肌、外套膜、鳃和肾等组织中的分布，定量RT-PCR和原位杂交技术揭示了Dmrt1 mRNA在性腺中的时空表达特征。结果显示：栉孔扇贝Dmrt1序列中含有DMRT家族保守的DM结构域；其mRNA仅在栉孔扇贝性腺中表达，在精巢中的表达明显高于卵巢，并且以生长期精巢的表达水平最高；原位杂交检测Dmrt1阳性信号主要定位于生殖细胞的细胞质中。研究结果表明，Dmrt1在栉孔扇贝性腺中的表达特征与大部分动物性腺中的表达特征基本一致，推测其可能参与性别分化和精巢发育的功能调节。