Quantity production of anther-derived haploids from a multiple disease resistant tobacco hybrid. I. Frequency of plants with resistance or susceptibility to tobacco mosaic virus (TMV), potato virus Y (PVY), and root knot (RK)

Summary Methods are described for producing large numbers of haploid plantlets from anthers of a flue-cured tobacco hybrid with monogenic resistance to tobacco mosaic virus, (TMV), potato virus Y (PVY) and root knot (RK), respectively. Additional details are given on colchicine treatment for converting haploids to doubled haploids (DH's) and on the frequency of spontaneous DH's among untreated plantlets. Disparate genetic ratios of TMV-resistant to TMV-susceptible plants were obtained among colchicine-treated haploid plantlets, induced DH's and untreated haploids when compared with F₂ and BC₁ progenies. Haploids (gametes) with the gene for TMV resistance occurred more frequently than expected and plantlets with the gene for RK resistance occurred less frequenctly than expected. Transmission of the gene for PVY resistance differed only slightly from Mendelian expectations. These unexpected ratios, in addition to the frequent occurrence of plastid chimeras among anther-derived plantlets, strengthened our conviction that haploidy is somehow associated with mutation.Joint contribution from the Departments of Genetics, Crop Science and Plant Pathology, North Carolina State University, and the Tobacco Research Laboratory, Agricultural Research, Science and Education Administration, U.S. Department of Agriculture, Oxford, North Carolina. Paper No. 5576 of the Journal Series of the North Carolina Agricultural Experiment Station, Raleigh, North Carolina.     

黄花蒿烟草醋液对三线镰刀菌生长的抑制作用

充分利用黄花蒿烟草秸秆制成秸秆混合醋液,并初步研究其对三线镰刀菌生长的影响。分离纯化真菌并对菌种进行传统形态学和分子学鉴定,利用干馏法制取黄花蒿烟草秸秆醋液,设置浓度梯度观察黄花蒿烟草秸秆醋液对三线镰刀菌2株菌的生长影响。鉴定获得的其中2株菌种均为三线镰刀菌,且在黄花蒿烟草秸秆醋液50%浓度下,生长抑制率的相对校正率近似为100%,达到完全被抑制的效果。研究结果为黄花蒿烟草秸秆潜在的利用价值研究及开发提供技术参考。     

烟草PR3b转录后剪切元件NRSE1与GUS融合表达后的可变剪切

【目的】烟草（Nicotiana tabacum L.）碱性几丁质酶基因PR3b在低烟碱突变体（nic1和nic2）中存在转录后mRNA可变剪切现象,但其可变剪切的发生机制仍不清楚。将PR3b的可变剪切元件NRSE1（nicotine- synthesis related splicing element 1）与GUS融合表达,分析NRSE1元件的独立可变剪切特性,以揭示其作用机制。【方法】利用PCR扩增方法获得PR3b cDNA序列中的NRSE1元件片段,并利用基因重组技术构建了烟草PR3b可变剪切元件NRSE1与GUS的融合表达载体。将融合表达载体导入农杆菌LBA4404后,通过农杆菌介导的叶盘转化法培育了表达NRSE1与GUS融合子的低烟碱突变体nic1和nic2及野生型烟草转基因植株;通过RT-PCR检测及GUS染色鉴定出阳性植株后,利用RT-PCR分析NRSE1与GUS融合表达后在低烟碱突变体和野生型烟草中的可变剪切特性;对转基因植株的幼苗进行乙烯（ET）和茉莉酸（JA）处理,通过GUS染色方法分析ET和JA处理对转基因植株中GUS活性的影响,并通过RT-PCR方法分析ET和JA处理对转基因植株中NRSE1与GUS融合子的可变剪切特性影响,以及对转基因植株中NRSE1与GUS融合子表达水平的影响。【结果】通过RT-PCR检测及GUS染色鉴定出表达NRSE1元件与GUS融合子的低烟碱突变体和野生型烟草转基因植株;RT-PCR检测及测序分析证明,NRSE1元件与GUS融合表达后仍能在低烟碱突变体发生高水平的可变剪切,剪切修饰区段的序列变化与烟草中PR3b的mRNA可变剪切修饰一致;利用ET和JA处理转基因植株进行的GUS染色表明,ET和JA处理对转基因植株的GUS活性有不同程度的影响;但利用ET和JA处理转基因植株进行的RT-PCR分析表明,ET和JA处理不改变NRSE1元件原有的诱导剪切特性,也不影响转基因植株中NRSE1元件与GUS融合子的表达水平。【结论】PR3b的可变剪切元件NRSE1与GUS在烟草中融合表达后,仍能在低烟碱突变体nic1和nic2中发生高水平的可变剪切;NRSE1在烟草中的可变剪切不依赖PR3b的其他mRNA区段,是烟草PR3b发生可变剪切的独立元件;ET和JA处理对NRSE1元件与GUS融合表达植株的GUS活性具有一定影响,可能存在翻译水平的调控作用。     

A key effector,BxSapB2, plays a role in the pathogenicity of the pine wood nematode Bursaphelenchus xylophilus

The pine wood nematode (PWN), Bursaphelenchus xylophilus, causes huge economic losses in pine forests. The plant‐parasitic nematodes have a complex life cycle that includes the secretion of effector proteins through a stylet into the host cell to promote parasitism. In this study, SignalP 4.1 and TMHMM 2.0 were used in preliminary screens for candidate effectors and were expressed in Nicotiana benthamiana through the PVX virus expression vector. The yeast signal sequence trap system was used to further study the function of the signal peptide of an effector, BxSapB2. In situ hybridization was conducted to investigate the localization of BxSapB2, followed by RNA interference technology (RNAi) to assess the functions of BxSapB2. The results demonstrate that BxSapB2 is a secreted protein that induces cell death in N. benthamiana and is highly expressed in esophageal gland cells and amphids of B. xylophilus. BxSapB2 was determined to be related to the pathogenicity of B. xylophilus. The results of this work indicate that BxSapB2 plays an important role in the interactions between B. xylophilus and the hosts.     

烟草(Nicotiana tabacum)非生物胁迫抗性分子机制研究进展

非生物胁迫影响烟草的生长发育,是制约作物产量和品质的重要因素。本综述阐述了非生物胁迫对烟草的影响以及最新的信号调控机制,包括干旱、盐、低温胁迫。在此基础上,综合分析了某些信号基因可能介导两因素甚至多因素调控过程,以及一些转录因子参与植物对冻害、干旱、盐害等非生物胁迫的应答调控,证明其对植物非生物胁迫的调控往往是多效性的。从分子水平上解析烟草在各种胁迫条件下的适应机制,对烟草种质资源保存、抗逆育种以及提高产量具有重要意义,为进一步研究植物的非生物胁迫应答提供理论依据。     

A heterogeneous potassium supply enhances the leaf potassium concentration of ridge‐cultivated tobacco grown in calcareous soil

The high potassium (K)‐fixing capacity of calcareous soils with homogeneous K supply limits crop K concentration. The objective of this study was to determine the increase in the leaf K concentration of tobacco (Nicotiana tabaccum L.) with a heterogeneous K supply. Two field experiments were conducted from 2013 to 2015 with five K rates applied in homogeneous environments and one K rate applied in varying fractions of soil volume (from 0% to 100%) in heterogeneous environments. Under ridge culture, leaf biomass was not significantly affected by homogeneous (K rate) or heterogeneous (patch size) K management. Under homogeneous conditions, increasing the K supply to three times the standard rate used by farmers (249 kg K ha⁻¹) increased leaf K concentration significantly, but the effect was limited and not cost‐effective. Heterogeneous supply of K fertilizer at the standard K rate in patches of 0–40% of soil volume outperformed homogeneous K supply in terms of leaf K concentrations. Furthermore, K fertilizer mixed in intermediate soil volume fractions of 1–20% resulted in higher leaf K concentrations than the smallest (0%) and larger (30–40%) soil fractions. However, such K‐rich patch application should be managed precisely with specific placement due to the sharp reduction in available K with increasing distance from the fertilizer. To maximize the sorption of immobile K, the match between the uptake volume of the rhizosphere and the area available for K diffusion is important.     

烟叶烘烤过程中多酚类物质的变化及与化学成分的相关分析

 研究了河南不同产烟区烟叶在烘烤过程中酚类物质含量变化及与化学成分的相关性。结果表明，总酚含量在烘烤开始后的前24 h呈增加趋势，之后则降低，至72 h降到最低，72 h后又迅速升高直到烘烤结束；绿原酸含量在烘烤0～24 h也呈上升趋势，其最低值出现在60 h或72 h处，之后则缓慢上升至烘烤结束；芸香苷含量随烘烤进程的推进，呈递增趋势，中间稍有起伏；多酚氧化酶（PPO）、过氧化物酶（POD）活性变化趋势相反。在烘烤过程中芸香苷的含量变化与总糖含量呈极显著正相关（r=0.822**），与淀粉、烟碱和蛋白质含量呈极显著负相关。     

湖南省烟草复混肥配方研究

继1986～1987年研制或A型肥和B型肥之后,1988年又根据湖南土壤,气候等条件,配制出6种复混肥(包括A型肥和B型肥),在24个植烟县(市)进行了肥效试验。结果表明,增施钾肥和施用稀土、硼、锌的综合效应能促进烟草生长发育,改善烟株农艺性状,显著或极显著地提高烤烟产量和质量.烤烟适宜施钾量为225～450kg/ha,平均为337.5 kg/ha;湘南、湘中烟区,施氮150～180kg/ha;湘西北烟区,施氮120～135kg/ha,能使烤烟达到优质适产的目标,1990年,选定氮:磷(P_2O_5):钾(K_2O)为10:12:24,加稀土、硼、锌和镁的配方生产出养分总含量达46%的“众望牌”烟草复混肥。