Development and application of a molecular marker for TMV resistance based on <Emphasis Type="Italic">N</Emphasis> gene in tobacco (<Emphasis Type="Italic">Nicotiana tabacum</Emphasis>)

Tobacco mosaic virus (TMV) caused serious loss in yield and quality of tobacco every year. It is a long-term goal to improve the tobacco resistance against TMV by tobacco breeding. N gene was the firstly reported TMV-resistant gene, which showed resistance against all Tobamoviruses except the Ob stain and belonged to the toll-interleukin-1 receptor/nucleotide-binding site/leucine-rich repeat class of plant resistance (R) genes. At present, N gene had already been widely used in tobacco conventional breeding, but there is rare available molecular maker used in marker-assisted selection of TMV resistance. In this study, we designed a pair of primers that specific amplify N gene fragment based on the sequence of N gene intron III, named N-marker. Then, we identified TMV resistance by two selecting methods, PCR with N-marker and inoculated with the TMV-C strain. Results from the two method showed that (1) 13 varieties among 67 tobacco varieties displayed hypersensitive reaction when inoculated with the TMV-C strain, also contained N gene fragments screened by PCR with N-marker; (2) 105 strains of 200 BC1 strains showed resistance against TMV when inoculated with TMV-C strain, meanwhile, 103 of the 105 strains contained N gene fragment verified by PCR with N-marker. Therefore, the N-marker is reliable for high throughput screening of germplasm resources and tobacco breeding materials in selection of N-mediated TMV resistance. Our study not only developed a molecular marker for tobacco breeding, but also identified new germplasm resources that are resistant to TMV.     

Two reported cytotypes of the emergent orchid model species <Emphasis Type="Italic">Erycina pusilla</Emphasis> are two different species

Each species is characterized by a specific set of chromosomes, which is described as the chromosome portrait or karyotype. In general, such a karyotype is the same for all individuals in the population. An exception to that rule has recently been found in the orchid Erycina pusilla, which has been reported to have two cytotypes with chromosome numbers of 2n = 10 and 2n = 12. Here, we examined the karyotypes of the two cytotypes and found differences in arm ratios and heterochromatin patterns as well as in the presence of satellite chromosomes and in the number and location of rDNA and telomeric repeat sites. These differences are extensive and would have required multiple chromosome rearrangements to generate the differences between the two karyotypes. We also found that F1 hybrids between the parents with the two different chromosome numbers resulted in sterile offspring, in accordance with our previous findings. The combination of hybrid sterility and extensively rearranged chromosomes supports the hypothesis that these two reported cytotypes are, in fact, two different species.     

簇毛麦6VS特异转录序列P21461及P33259的获得及其分子标记在鉴定小麦-簇毛麦抗白粉病育种材料中的应用

簇毛麦6V#2S和6V#4S染色体臂分别携带抗白粉病基因Pm21和Pm V,在与小麦的杂种后代中,抗病基因与外源染色体臂共分离。开发鉴定2条外源染色体臂间多态性的序列,尤其是遗传信息相对缺乏的6V#4S染色体臂的序列,对于其在遗传与育种上的应用具有重要意义。本研究以携带6V#4S·6DL染色体的小麦易位系Pm97033及感病小麦亲本宛7107接种白粉菌的叶片转录组数据为资源,通过差异基因筛选、共线性分析、簇毛麦基因组扩增及测序验证的方法,鉴定出来自6V#4S的表达序列P21461和P33259,其中基于P21461序列设计的引物P461-5在簇毛麦6V#2S和6V#4S染色体臂的扩增产物具有30 bp的In Del和4 nt的多态性。用该引物转化的标记P461-5a可以鉴定抗白粉病小麦品种和高代品系所含的外源染色体,显示其在簇毛麦抗源鉴别和小麦抗病育种辅助选择中潜在的应用价值。根据P33259开发的标记P259-1可以对含有6V#4S染色体臂的材料进行特异扩增,但对6V#2S·6AL易位染色体没有扩增产物,因此P259-1可作为6V#4S·6DL易位染色体的特异分子标记。q RT-PCR分析结果显示,P21461的表达不受白粉菌诱导,而P33259在接菌后12 h和24 h的转录水平比接菌前提高约2倍,推测其可能参与Pm97033与白粉菌的早期互作。     

基于LiDAR的亚热带次生林林窗对幼树更新影响分析

以湖南亚热带次生林为研究对象,利用多时相机载激光雷达(Light detection and ranging,Li DAR)和野外调查数据对林窗及幼树进行监测,分析比较林窗对幼树密度分布和树高生长变化的影响。结果表明,林窗大小和位置对幼树密度分布都有显著影响,喜光树种幼树主要集中在小林窗的中心区或大林窗的过渡区,在大林窗中密度最大(647株/hm2),耐荫树种幼树主要集中在林窗的边缘区,在中等林窗中密度最大(941株/hm~2)。林窗大小对幼树树高生长有显著影响,喜光树种和耐荫树种幼树分别在大林窗和中等林窗中树高生长量最大(69.3 cm/a、57.7 cm/a),喜光树种幼树在中心区的树高生长量明显大于其他位置,耐荫树种幼树的树高生长量在位置上的差异不显著。线性混合模型分析显示林窗大小是促进幼树树高生长的最主要因素,幼树树高生长变化在不同林窗中呈聚集性。从幼树密度树高生长情况来看,50~150 m~2林窗较适合促进亚热带次生林的群落演替。     

豌豆根腐病研究进展

根腐病是豌豆根部的重要病害之一,在世界各地豌豆产区均有发生,是制约豌豆产业持续健康发展的因素之一。世界上尚未发现对根腐病完全免疫的豌豆品种,防治方法主要以农业防治和化学防治为主。本文从豌豆根腐病的发生与分布、病原菌的分类及特点、抗性鉴定及评价标准、种质资源、分子标记及防治策略等方面对国内外豌豆根腐病研究现状进行综述。并提出抗病育种和未来豌豆根腐病综合防治的研究方向。     

四种菊酯类杀虫剂对枸杞蚜虫线粒体膜流动性的影响

为明确菊酯类杀虫剂对昆虫线粒体膜流动性的影响,以1,6-二苯基-1,3,5-己三烯（1,6-diphenyl-1,3,5-hexatriene,DPH）为荧光探剂,采用荧光偏振法研究了4种菊酯类杀虫剂对枸杞蚜虫Aphis sp.线粒体膜流动性的影响。结果表明,4种菊酯类杀虫剂对枸杞蚜虫线粒体膜流动性均有显著影响,其荧光偏振度显著增大,线粒体膜流动性显著减小。当药剂浓度为100 μmol/L时,对照组枸杞蚜虫线粒体膜的荧光偏振度为0.239,高效氯氰菊酯的影响最大,线粒体膜的荧光偏振度为0.315,联苯菊酯的影响最小,线粒体膜的荧光偏振度为0.287。4种菊酯类杀虫剂对线粒体膜流动性的影响都存在一定的浓度效应,当药剂浓度在0~100 μmol/L范围内,浓度越大影响越大。4种菊酯类杀虫剂在不同温度下对枸杞蚜虫线粒体膜流动性的影响不同,高效氯氰菊酯、联苯菊酯和甲氰菊酯对线粒体膜流动性的影响基本不受温度影响,而氯菊酯在高温条件下产生的影响大,在低温条件下产生的影响小,表现为明显的正温度系数。表明DPH可以作为荧光探剂研究枸杞蚜虫线粒体膜的流动性。     

花期低温对水稻叶鞘生理及结实率的影响

盆栽条件下,以耐冷性不同的2个水稻（Oryza sativa L.）品种龙稻5（耐冷型）和龙粳11（冷敏型）为材料,于开花期在人工气候室进行低温（15℃,分别持续1,2,3,4,5 d）处理,研究开花期低温对不同耐冷性水稻结实率及叶鞘膜透性、抗氧化酶等生理指标的影响。结果表明：低温处理2 d时,龙稻5和龙粳11的花粉活力分别下降了8.21%和16.10%,差异达极显著水平;低温处理3 d后,龙稻5的结实率下降到99.90%,与CK相比差异达到显著水平;低温处理1 d后,龙粳11的结实率下降到60.04%,与对照相比差异达到极显著水平。低温处理提高了水稻叶鞘相对电导率和脯氨酸、可溶性蛋白含量,处理5 d时各指标达最大值,与CK相比,龙稻15和龙粳11分别提高了38.01%和20.77%、25.06%和84.85%、25.82%和23.63%。同时低温处理也提高了叶鞘超氧化物歧化酶（SOD）、过氧化物酶（POD）和过氧化氢酶（CAT）活性,处理3 d时酶活性达最大值,龙稻15和龙粳11分别较CK提高了10.57%和14.90%、86.26%和68.71%、17.73%和28.25%。综上所述,开花期低温引起水稻叶鞘膜脂过氧化程度升高,但也诱导水稻叶鞘抗氧化系统开启、保护酶活性上升以抵御低温伤害;开花期低温导致水稻花粉活力降低并引起结实率降低,耐冷品种较冷敏品种抵御能力强。     

"沃得天"微肥叶面喷施对猕猴桃产量和品质的影响

验证"沃得天"微肥(一种新型微量元素水溶肥)对猕猴桃果树的喷施肥效,为陕西省猕猴桃生产提质增效提供科学依据。试验设置了喷施新型微量元素水溶肥("沃得天"微肥1(W1)、"沃得天"微肥2(W2))、普通微量元素水溶肥(G1)、清水(当地地下水,CK1)4个处理,供试材料为"秦美"猕猴桃,田间采用完全随机区组设计,测定了叶片生长、果实产量和品质方面的指标,并综合评价了"沃得天"微肥的田间应用效果。结果表明,与对照相比,喷施W1和W2均显著提高了猕猴桃叶片的百叶重和百叶厚,百叶重分别增加22.6%和33.3%,百叶厚分别增加20%和27%;其中W2还能同时增加果实横、纵径,增幅为11.0%和12.9%,并能够显著提高叶片中Cu、Zn、Fe、Mn以及Ca的含量,其中与普通微肥相比增加Fe元素60～78 mg·kg~(-1)。喷施W1和W2分别增加猕猴桃单果重为11.3%和8.8%,增加了果实的固酸比,其中W2果实中Vc的含量显著增加了8.6%;W1和W2还不同程度增加了果实中多种微量元素和钙的含量,优化了猕猴桃果实外观和营养品质。另外,W1和W2处理猕猴桃分别增产8.11%和5.17%,增益8.83%和5.43%,而且2种"沃得天"新型微量元素肥料对猕猴桃增产提质效果均优于市售普通微肥。因此,叶面喷施"沃得天"新型微肥可以促进陕西"秦美"猕猴桃的生长发育,并且能在一定程度上调控叶片和果实对必需微量元素的累积,在提高果实品质和增产增收及优化猕猴桃肥料管理方面效果显著。     

Effect of CHOP expression knock-down on apoptosis of renal tubular epithelial cells

AIM:To study the effect of C/EBP homologous protein (CHOP) on the apoptosis of renal tubular epithelial HK2 cells. METHODS:The serum mRNA levels of CHOP in the patients with acute kidney injury and healthy controls were detected by qPCR. In vitro, renal tubular epithelial HK2 cells were divided into control group, negative group (transfected with negative control siRNA), si-CHOP group (transfected with CHOP siRNA), and induced by transforming growth factor-β1 (TGF-β1). The viability of the cells was measured by MTT assay, and the apoptotic rate was analyzed by flow cytometry. The protein levels of nuclear antigen Ki-67, proliferating cell nuclear antigen (PCNA), caspase-3 and cleaved caspase-3 were determined by Western blot. RESULTS:Compared with the healthy controls, the serum mRNA levels of CHOP in the patients with acute kidney injury were increased significantly (P<0.05). Transfection with CHOP siRNA significantly decreased the expression of CHOP in the renal tubular epithelial HK2 cells (P<0.05). Knock-down of CHOP expression by siRNA significantly increased the viability of renal tubular epithelial HK2 cells (P<0.05), decreased the apoptotic rate (P<0.05), increased the expression of Ki-67 and PCNA (P<0.05), and down-regulated the protein level of cleaved caspase-3 (P<0.05). CONCLUSION:The serum mRNA levels of CHOP were increased in the patients with acute kidney injury. Knock-down of CHOP expression inhibits the apoptosis of renal tubular epithelial cells by regulating the expression of proliferation-and apoptosis-related proteins.     

Effects of histone hypomethylation induced by alcohol during pregnancy on overexpression of cardiomyogenesis genes offspring mice

AIM:To investigate the effects of histone methylation on the abnormal expression of cardiomyogenesis genes caused by alcohol during pregnancy and the regulatory mechanism, and to provide a new idea and intervention targets for preventing and curing congenital heart disease. METHODS:The alcohol (56%, 5 mL/kg) and G9a-histone methyltransferases (HMT) inhibitor BRD4770 (1 mg/kg) were given by gavage in Kunming mice during embryo (E) 0.5~14.5 d, and the hearts of the mice in E14.5, E16.5 and post neonatal 0.5 d (PND0.5) were collected. The mRNA expression of Gata4, Cx43 and β-MHC genes was detected by RT-qPCR. The activity of HMT was measured by colorimetry. Meanwhile, the protein expression of histone H3K9me3, G9a-HMT, Cx43 and β-MHC was determined by Western blot. RESULTS:The results of colorimetry showed that the activity of HMT in the heart of the offspring mice treated with alcohol during pregnancy was decreased significantly compared with normal saline group (P<0.05), and Western blot data showed that the expression of G9a-HMT and histone H3K9me3 were apparently decreased in the same samples (P<0.05). The mRNA expression levels of Gata4, Cx43 and β-MHC in alcohol group were apparently increased compared with normal saline group (P<0.05). Meanwhile, the protein levels of Cx43 and β-MHC were increased significantly in the same samples (P<0.05). However, BRD4770, a G9a-HMT inhibitor, further attenuated the level of histone H3K9me3, and further upregulated the expression of Gata4, Cx43 and β-MHC in the heart of the the mice treated with alcohol (P<0.05). CONCLUSION:Histone methylation modification imbalance induced by G9a-HMT may be involved in the abnormal expression of cardiomyogenesis genes in the heart of offspring mice caused by alcohol during pregnancy.