马铃薯耐盐胁迫品种(系)的初步鉴定与筛选

为了筛选出适合盐碱地种植的马铃薯耐盐品种（系）,本试验以来自不同地区的6 个马铃薯品种（系）为材料,在植物组织培养条件下,用不同浓度的NaCl模拟盐胁迫环境,通过比较盐胁迫对植株外观性状和生理特性的影响,对耐盐品（系）进行初步鉴定与筛选。结果表明：‘东农09-33069’在成活率、生根率、生根数、根长、叶片数、株高、茎粗等形态指标和叶绿素相对含量、根系活力、脯氨酸含量等生理指标上均表现优于其他5个参试品种（系）,且其他5个品种（系）均在部分指标上表现出对盐胁迫的耐受性较差。因此,在6 个参试品种（系）中‘东农09-33069’的盐胁迫耐受性强、稳定性高,可初步鉴定为耐盐品种（系）。     

亲水短肽基因NLEAs提高酵母的耐盐能力

胚胎晚期富集蛋白(late embriogenesis abundant protein, LEA)是植物种子成熟过程中富集的蛋白质,与植物应答逆境胁迫密切相关,具有很强的亲水性。LEA蛋白的增多,有利于降低细胞水势,增强植物耐盐胁迫等逆境的能力。为了进一步提高LEA蛋白的亲水性,提高细胞耐受盐胁迫等渗透胁迫的能力,本研究对LEA蛋白数据库进行了生物信息学分析。结果鉴定出一系列亲水性高的氨基酸序列。对这些序列进行重复及随机组合后,合成了假定的NLEA (Novel LEA)基因。通过筛选转基因酵母菌株,获得了有助于提高酵母耐盐性的NLEA8和NLEA13基因。同时,初步分析了NLEA8和NLEA13基因表达的蛋白的与植物耐盐相关的生物学功能和结构特点。本试验获得了LEA基因,通过酵母耐盐性筛选验证其耐盐能力,对耐盐基因的获得及验证提供了一定的参考。     

水稻耐盐性的遗传和分子育种的研究进展

盐胁迫是造成水稻减产的重要环境因素之一。本文从维护膜系统的完整性、离子的区隔化以及渗透调节等三个方面介绍了水稻耐盐性的机理;简要描述了水稻生物耐盐能力、农艺耐盐能力和离体细胞对盐害反应等三种耐盐鉴定方法。总结了近年来对水稻耐盐种质资源的发掘、耐盐性QTL定位和重要基因的克隆以及耐盐水稻选育所取得的进展。通过长期水稻耐盐性评价、已发掘了一些耐盐的水稻种质;定位了七十多个控制Na+/K+含量、存活天数等耐盐性相关性状的QTL;两个水稻耐盐基因SKC1和DST已被克隆;我们已获得系列不同程度耐盐的转基因植株和SKC1/BADH两类耐盐基因聚合系。本文进一步讨论了水稻耐盐性机制的研究以及在生产实践中应用的前景,试图为深入开展水稻耐盐性研究提供参考。     

复垦滩涂微咸水灌溉下粳稻产量和品质的表现

为明确不同粳稻品种在江苏沿海滩涂地区的产量和品质表现,以近期育成的23个新品种（系）为材料、其中盐稻12号为对照,在中低盐分（2g/kg）复垦滩涂地块,微咸水（矿化度0.94~2.44g/L）灌溉,比较滩涂实地盐胁迫下粳稻产量和品质的表现。结果表明,参试品种（系）平均产量8.21t/hm²,变幅6.19~11.74t/hm²,为邻近无盐胁迫老垦区产量的85%左右;主要的穗粒结构表现为穗多而粒少,增加穗粒数是滩涂水稻高产栽培途径之一。就稻米品质而言,滩涂中低度盐胁迫对糙米率、精米率、长宽比和食味值的影响不大,对整精米率有一定的影响,影响最大的是稻米外观品质指标垩白粒率和垩白度,在耐盐品种选育与筛选中要加强对垩白度的选择。     

水稻籼型不育系萌发期和幼苗期的耐盐性评价

为了鉴定水稻籼型不育系在不同生育阶段的耐盐性,筛选出适合耐盐杂交水稻新组合测配的强耐盐不育系,笔者分别在萌发期、幼苗期对8个水稻籼型不育系进行耐盐性鉴定,以耐盐品种FL478、盐敏感品种IR29为对照。结果显示：盐胁迫对水稻种子的萌发起到抑制作用,抑制效应随盐浓度的提高而增强,10 g/L NaCl浓度胁迫能较好反映水稻不育系萌发期耐盐性的差异。‘沪旱7A’、Y58S在萌发期的耐盐性最好,与FL478均属于1级,具极强的耐盐性;‘荃9311A’的盐害级别为3级,具强耐盐性。盐胁迫对水稻幼苗的生长也起到明显的抑制作用,8 g/L NaCl浓度胁迫下的枯叶率性状能较好反映水稻不育系幼苗期耐盐性的差异,‘沪旱11A’与耐盐对照FL478耐盐性相仿。     

转GHABF4转录因子棉花植株的耐盐性研究

AREB/ABFs转录因子家族基因主要参与干旱、高盐、低温等胁迫应答反应。为了获得具有较高耐盐水平的棉花新种质材料,通过农杆菌介导法将耐盐转录因子基因(GHABF4)导入陆地棉中棉35中,通过对转化植株的卡那霉素初步筛选及T1、T2、T3目的基因PCR的分子检测,获得T3转基因棉花纯合系。通过盐胁迫试验对5个T3转基因棉花株系和非转基因棉花对照进行耐盐性分析。结果表明,在200 mmol/L Na Cl胁迫下,与非转基因对照相比,5个转基因棉花株系株高提高2.5~4.4 cm,地上部分的鲜质量增加3.6%~11.8%;且抗氧化物酶SOD、POD、CAT活性以及叶绿素含量提高。在盐胁迫条件下,转GHABF4基因棉花表现出优良的生长和生理优势,转GHABF4基因能够提高棉花的抗盐能力。     

甘蓝型油菜盐胁迫下幼苗鲜重和干重QTL定位及候选基因分析

盐胁迫是非生物胁迫中影响作物产量的一个主要因素,利用分子标记方法选育油菜耐盐品种对提高油菜产量具有重要意义。选用来自GH06与P174杂交后通过单粒传法连续自交获得的高世代重组自交系群体,以含16 g L–1Na Cl的Hoagland溶液培养幼苗进行盐胁迫处理25 d后,分别测定叶和根的鲜重及干重,根据已构建的高密度SNP遗传连锁图谱进行QTL定位,在QTL物理区间筛选耐盐相关基因并以极端表型材料进行q RT-PCR分析。采用复合区间作图法(CIM),在对照和盐胁迫处理中共检测到19个QTL,其中与盐胁迫相关的有6个,可解释的表型变异7.16%~16.15%,分布在A02、A04和C03染色体上,将QTL置信区间序列和拟南芥中与盐胁迫相关的基因比对分析,共找到8个候选基因。对其中4个候选基因在极端表型材料中的表达分析表明,BnaA02g14680D与BnaA02g14490D基因在盐胁迫处理后的48 h或72 h表达量均高于对照组,即基因的表达由盐胁迫引起,而BnaC03g64030D在敏感型材料中的相对表达量高于在耐盐型材料中,BnaC03g62830D在敏感型材料中没有明显变化,但在耐盐型材料中呈现先升高后降低的表达特征,其表达可能会增强植株对盐胁迫的耐受力。本研究为油菜耐盐基因功能挖掘和油菜耐盐品种选育奠定基础。     

蒙古柳cDNA转化酵母菌及NaCl胁迫抗性基因筛选

为挖掘蒙古柳的NaCl胁迫相关基因,本研究利用FOX hunting system技术,构建蒙古柳cDNA连接pYES2载体后转入酵母InVSCI菌株。本研究将pYES2-蒙古柳cDNA酵母菌在1.0 mol/L Na 抗性板胁迫处理后得到15个抗NaCl胁迫的酵母菌株,并利用这15个菌株进一步通过NaHCO3、H2O2和Srobitol抗性进行分析,结果发现都表现出比对照组更强的耐胁迫性。接着对这15个菌株提取细胞DNA后克隆获得蒙古柳cDNA,在NCBI数据库中对测序得到的基因序列进行相似性比对,结果发现大部分和毛果杨基因有高的相似性,其中一些基因已有研究表明和盐胁迫相关。本研究可为挖掘可利用的耐盐基因资源提供依据。     

不同基因型花生抗旱耐盐性评价及鉴定指标筛选

旱盐交叉加剧了盐胁迫对花生的危害,评价鉴选抗旱耐盐品种对于盐碱地花生生产具有重要意义。本研究采用盆栽试验,设置0.3%盐胁迫和中度干旱胁迫,对30个花生品种的出苗率、植株形态和生物量等指标进行抗旱耐盐系统评价。结果表明,发芽率和生物量可作为抗旱耐盐评价的首选指标,分枝数、第一侧枝长可作为辅助指标。旱盐胁迫下,30个品种可分成高度抗旱耐盐型、抗旱耐盐型、旱盐敏感型和高度旱盐敏感型。其中高度抗旱耐盐型5个,占供试品种的16.67%;抗旱耐盐型13个,占供试品种的43.33%;旱盐敏感型为9个,占供试品种的30%;高度旱盐敏感型3个,占供试品种的10%。本试验为花生抗旱耐盐机理研究及生产应用提供了不同类型材料。     

NaCl 胁迫下不同品种（系）绿豆种子萌发期耐盐性的研究

为了探究盐胁迫对不同品种（系）绿豆的影响,对 28 份绿豆种子用 1.2%NaCl 进行盐胁迫处理,测定了与萌发相关的 12 项指标,采用相关性分析、主成分分析、隶属函数分析及聚类分析对不同品种（系）进行了综合评价和分类。1.2%NaCl 胁迫对不同绿豆品种（系）12 项测定指标有显著影响,各指标间存在着不同程度的相关性。主成分分析结果表明,前 3 个主成分累计贡献率为 88.072%;抗旱综合评价 D 值各不相同,其中同绿 9 号的抗旱综合评价值最大（0.681）,表明其对盐胁迫的耐受性最强,168-21 的抗旱综合评价值最小（0.444）,表明其对盐胁迫的耐受性最弱。根据不同品种（系）绿豆耐盐性隶属函数 D 值进行了聚类分析,共分为 3 类：第Ⅰ类为耐盐品种（系）,第Ⅱ类为中等耐盐品种（系）,第Ⅲ类为耐盐性较弱品种（系）。综合评价各项指标,同绿 9 号耐盐性最强,168-21 耐盐性最弱,可为盐碱地种植绿豆提供参考。     

In vitro mutagenesis and directed screening for salt-tolerant mutants in peanut

To expand the salt-tolerant gene resources of peanut, we conducted in vitro mutagenesis with pingyangmycin (PYM) as the mutagen and directed screening with medium containing NaCl. After embryonic leaflets from mature peanut seeds (variety Huayu 22) were cultured on somatic embryogenesis-induction medium containing 4 mg/L PYM for 4 weeks, the surviving somatic embryos were sequentially transferred to a germination medium containing 15 and then 20 g/L NaCl. The 30 NaCl-tolerant plantlets obtained were grafted and transplanted in the field in 2011, and the mature seeds of 26 regenerated plants were harvested. In 2012, all seeds from each plant were sown in the field. The offspring (M₂ generation) of 23 of 26 NaCl-tolerant, regenerated plants differed from their mutagenic parent in vigor, growth habit, flowering habit, pod shape, and seed coat color, and they also exhibited trait segregation from the same NaCl-tolerant, regenerated plant. In a germination test with a 0.7 % NaCl solution and the M₃-generation seeds from 18 of the NaCl-tolerant, regenerated plants, the germination rate was substantially higher for the seeds from 6 plants than for seeds from the mutagenic parent (Huayu 22). To determine whether the changes in plant traits might be associated with gene mutations, DNA polymorphisms between the mutagenic parent and 19 M₃ generation individuals from different NaCl-tolerant, regenerated plants were analyzed with 39 pairs of SSR primers, and all mutants differed from the mutagenic parent in >2 loci. The results indicate that the use of PYM-based mutagenesis in combination with directed in vitro screening with NaCl is effective for creating and identifying salt-tolerant mutants of peanut.     

临界生长温度锻炼对棉苗抗冷性的影响

研究了棉苗经过不同低温(12℃、14℃、16℃)锻炼7 d后,对较长时间(7 d)的低温(4℃)胁迫的耐受能力,并探讨了与其相关的可能机理.结果表明:经不同低温锻炼7 d后,所有棉花品种的幼苗脯氨酸含量都有不同程度的增加,其中12℃临界生长温度锻炼后增幅最明显;临界生长温度(12℃)下锻炼后的棉苗能耐受较长时间(7 d...     

农杆菌介导轮状病毒抗原蛋白VP4基因遗传转化花生的研究

选用鲁花14与花育23花生品种, 通过农杆菌介导开展了轮状病毒抗原蛋白VP4基因遗传转化研究, 从转化植株中随机选取26株表现Kan抗性植株进行npt II基因的PCR检测, 结果有22株能扩增出620 bp左右的npt II基因条带, 阳性率约为84.62%。提取npt II基因显示为阳性的植株叶片DNA作模板, 用VP4基因特异引物进行PCR扩增, 经琼脂糖凝胶电泳分析, 所有npt II基因阳性的植株均扩增出了约2 350 bp的特异性条带, 而野生型没有。对部分转基因植株进一步进行PCR-Southern杂交和Southern杂交分析, 发现转基因植株中出现了阳性杂交信号, 表明VP4基因的确已经整合到花生的基因组中, 并且是1~2个拷贝。用RT-PCR分析了11株转G1VP4基因的植株, 证明插入鲁花14中的VP4基因已经正常转录, 利用Western blot方法检测筛选到的4株转基因花生, 分别提取其蛋白, 在30 kD处出现特异性蛋白条带, 这为获得转基因创新种质材料奠定了基础。     

CPPU对干旱胁迫下小麦幼苗叶片的蛋白响应及生育期差异的影响

采用叶面喷施调吡脲(N-(2-chloro-4-pyridyl)-N’-phenylurea,CPPU)处理,分析干旱胁迫下CPPU对小麦幼苗叶片蛋白质及各种生理生化指标的影响,以探讨CPPU提高小麦抗旱性的分子机制。以小麦品种河农822为材料,用聚丙烯酰胺凝胶电泳法(SDS-PAGE)研究喷施CPPU后幼苗叶片中蛋白质组变化情况;用常规方法检测过氧化物酶(POD)活性、可溶性蛋白、可溶性糖、脯氨酸和丙二醛(MDA)含量的变化。结果表明,经SDS-PAGE电泳技术发现,与对照相比,干旱胁迫下喷施CPPU后出现分子量为125,80,55,40,35,33,25 kDa等7条特异性条带,消失了30 kDa的电泳条带。与干旱对照相比,喷施CPPU后叶片中蛋白条带中多了40 kDa特异性条带,消失了30 kDa的电泳条带;并且在干旱胁迫下,喷施100 mg CPPU后使幼苗体内的POD活性提高37.06%,可溶性蛋白含量增加63.50%,可溶性糖含量增加87.51%,脯氨酸含量增加109.57%,MDA含量降低46.06%。与干旱对照相比,喷施CPPU的幼苗叶片中出现了一条特异性条带,消失了一条特异性条带,且在70 kDa和大约20 kDa处的蛋白含量明显高于2个对照,在32kDa和大约25 kDa处的蛋白含量下降。100 mg CPPU可促进干旱胁迫下幼苗体内的可溶性糖、可溶性蛋白和脯氨酸含量显著增加,POD活性显著提高,MDA含量降低。说明叶面喷施CPPU能够增强幼苗对干旱胁迫的适应性。     

Baking quality of hard winter wheat: Response of cultivars to environment in the Great Plains

Inconsistency of wheat end-use quality has long been a problem for the milling and baking industries, which require high levels of uniformity for modern, high speed, processing. Extensive research was conducted to characterise genetic, environmental, and biochemical factors that contribute to variation in wheat quality. Samples from 17 locations of trials with 30 cultivars grown over two years were evaluated for grain quality, milling yield, mixograph parameters and baking properties. Protein quality and composition of flour samples were determined by SDS sedimentation and size-exclusion chromatography. Test sites were monitored for soil fertility, and meteorological data was collected hourly during grain fill. Additional grain samples and data were collected from six cultivars grown at 11 locations in 1993. Significant variation in end-use quality was observed among samples; variation was attributed to environmental effects, genotype, and their interactions. For many mixograph and baking parameters, variation attributed to environmental effects was of greater magnitude than for genotype. Increasing levels of low molecular weight (LMW) saline soluble proteins were related to a general decline in dough mixing properties and loaf characteristics. However, correlations of protein components with baking parameters were generally low, suggesting limited predictive value. Hours of high temperature stress (>32 °C) during grain fill were associated with loaf volume and SDS sedimentation volume, although in a curvilinear fashion. Increasing temperature stress initially had a positive influence on loaf volume and protein quality; however, when temperature stress exceeded 90 hours, there was a strong negative influence on baking quality. Variation among cultivars for environmental stability was indicated and cultivar responses to temperature stress varied. Relationships with meteorological data suggest opportunities to predict wheat end-use quality through environmental modelling. This revised version was published online in August 2006 with corrections to the Cover Date.     

农杆菌介导轮状病毒抗原蛋白VP4基因遗传转化花生的研究

选用鲁花14与花育23花生品种, 通过农杆菌介导开展了轮状病毒抗原蛋白VP4基因遗传转化研究, 从转化植株中随机选取26株表现Kan抗性植株进行npt II基因的PCR检测, 结果有22株能扩增出620 bp左右的npt II基因条带, 阳性率约为84.62%。提取npt II基因显示为阳性的植株叶片DNA作模板, 用VP4基因特异引物进行PCR扩增, 经琼脂糖凝胶电泳分析, 所有npt II基因阳性的植株均扩增出了约2 350 bp的特异性条带, 而野生型没有。对部分转基因植株进一步进行PCR-Southern杂交和Southern杂交分析, 发现转基因植株中出现了阳性杂交信号, 表明VP4基因的确已经整合到花生的基因组中, 并且是1~2个拷贝。用RT-PCR分析了11株转G1VP4基因的植株, 证明插入鲁花14中的VP4基因已经正常转录, 利用Western blot方法检测筛选到的4株转基因花生, 分别提取其蛋白, 在30 kD处出现特异性蛋白条带, 这为获得转基因创新种质材料奠定了基础。     

Increased regeneration from NaCl-tolerant callus in rice

Summary NaCl-tolerant calli were selected from two Japonica and two Indica rice (Oryza sativa L.) cultivars on basal media containing 6,000, 9,000, 12,000 or 15,000 ppm NaCl. Frequency of callus formation decreased with the increase of NaCl in the medium, especially in Indica. About half of the calli of Japonica cultivars selected on NaCl-ammended media survived 20,000 ppm NaCl but none of the Indica callus survived. In Japonica, more plants were regenerated from calli selected on all concentrations of NaCl media than from NaCl-free medium. Concentration of Cl^- in callus increased dramatically with increased NaCl content but peroxidase activity decreased.     

小麦质不育系花药及雌蕊邻苯二酚氧化酶同工酶分析

小麦细胞质雄性不育系、保持系花药三个不同发育时期的邻苯二酚氧化酶同工酶存在明显差异,不育系中的同工酶谱带数多于保持系.考虑到小麦细胞质雄性不育系的细胞核与保持系相同,但具有与保持系不相同的不育细胞质,故邻苯二酚氧化酶同工酶可能与植物的雄性不育特性有某种联系.雌蕊中的邻苯二酚氧化酶同工酶在不育系与保持系间基本相同,这可能与雌蕊中不育系、保持系间的核质组成相同有关,这一事实似乎从另一侧面说明了邻苯二酚氧化酶与不育特性存在着某种联系.     

In vitro selection of wheat callus tolerant to high levels of salt and plant regeneration

Summary Embryogenic calli isolated from immature embryos of four wheat cultivars were subjected to three in vitro selection methods for salt tolerance. The effect of NaCl on the selected and unselected cell lines has been investigated. The results indicated that the relative growth rate of callus decreased as the concentration of NaCl increased in both callus lines. The selected callus line gave a higher growth weight in the presence of NaCl in the medium and was highly significant as compared with unselected callus line across medium protocols in all wheat cultivars. The dry weight of both kinds of callus lines of all wheat cultivars increased markedly with increasing NaCl concentration in most cases. The Na⁺ and Cl^- contents of both callus lines were increased with increasing salinity levels while K⁺ content was decreased. The selected callus line of each cultivar at the same salinity level produced significant amounts of Na⁺, K⁺ and Cl^- higher than the unselected callus line in most salinity levels. However, the unselected callus lines of the cultivars Giza-157 and Sakha-90 at the same salinity level produced significant amounts of K⁺ higher than the selected callus line in most salinity levels. The proline content of both kinds of callus lines for all wheat cultivars was increased with increasing salinity level. However, the selected callus line gave a significantly higher proline content than the unselected callus line in all wheat cultivars at the same Salinity level. Results from the in vitro selection for NaCl tolerance showed that the stepwise method of increasing NaCl in the medium was more effective for plant regeneration than other methods.     

盐胁迫对冬小麦过氧化物酶同工酶的影响

利用聚丙烯酰胺凝胶垂直板电泳法,研究了不同浓度ＮａＣｌ对冬小麦根（三叶期）过氧化物酶（ＰＯＤ）同工酶的影响,电泳结果显示：１ｍｇ／ｇ ＮａＣｌ胁迫电泳图谱Ａ区酶带较对照组增加２条,Ｂ区酶带增加１～３条;２ｍｇ／ｇ ＮａＣｌ胁迫时,Ａ区为１条酶带,Ｂ区酶带增加了３条;ｔ检验表明：１ｍｇ／ｇ ＮａＣｌ胁迫时Ｂ区新增３条酶带的品种的株高和穗长极显著（Ｐ〈０．０１）高于Ｂ区新增１条酶带的品种。