EMS Induction of Early Flowering Mutants in Spring Rape (Brassica napus)

Studies were undertaken to induce early flowering mutants by ethyl methanesulphonate (EMS) treatments of Brassica napus seeds. EMS treatments for 12 h of a highly inbred B. napus line TBS had adverse effects on M¹ plant development and fertility only when concentrations were greater than 1%. However, an EMS concentration of 1.5% did not reduce M₁ plant fertility to an extent which significantly reduced production of M₂ seeds. Genetic changes induced by EMS treatment and affecting flowering time were of three main types: (1) Changes within a polygenic system reflected by increased variation in flowering time among M₂ families. As the increase in variation was due primarily to a higher frequency of later flowering plants, these polygenic changes would be of little value in developing better-adapted cultivars. (2) Induction of a recessive mutation at a major gene locus which caused M₃ plants homozygous for the mutant gene to flower at least 20 days earlier than the parental line TB8. (3) Induction of a dominant mutation at a major gene locus which affects flowering time by causing a substantial reduction in vernalization requirement. M₂ plants carrying the mutant gene flowered as early as 59 days before the parental line. These major gene mutations could be rapidly exploited in the development of agronomically superior cultivars for short-season, lower rainfall environments in Western Australia.     

Broad-few-leaflets and outwardly curved wings: two new mutants of chickpea

This study was aimed at the induction of morphological mutations for increasing genetic variability and making available additional genetic markers for linkage studies in chickpea (Cicer arietinum L.). A wilt‐resistant, well‐adapted chickpea cultivar of central India,‘JG 315’(Jawahar gram 315), was used for the induction of mutations. Seeds presoaked in distilled water for 2 h were treated with ethyl methane sulphonate (EMS) using six different concentrations (0.1, 0.2, 0.3, 0.4, 0.5 and 0.6%) and two different durations (6 and 8 h). Several morphological mutants were identified in M₂. One of the mutants, isolated from a treatment of 0.3% EMS for 8 h, had five to nine large leaflets per leaf in comparison with 11‐17 normal‐sized leaflets per leaf observed in the parental cultivar ‘JG 315′. The mutant was designated broad‐few‐leaflets. Many leaves of this mutant showed a cluster of three to five overlapping leaflets at the terminal end. The other mutant, designated outwardly curved wings, was isolated from the 0.5% EMS treatment for 6 h. In this mutant, the wings were curved outwards, exposing the keel petal, while the wings in typical chickpea flowers are incurved and enclose the keel. The lines developed from the broad‐few‐leaflets and outwardly curved wings mutants were named JGM 4 (Jawahar gram mutant 4) and JGM 5, respectively. Inheritance studies indicated that each of these mutant traits is governed by a single recessive gene. The gene for broad‐few‐leaflets was designated bfl and the gene for outwardly curved wings was designated ocw. The locus bfl was found to be linked with the locus lg (light green foliage) with a map distance of 18.7 ± 6.3 cM.     

EMS诱导小麦TcLr19感叶锈病突变体的筛选

为了利用小麦感叶锈病突变体进行抗病基因功能和机制研究,用EMS处理小麦抗叶锈病近等基因系Tc Lr19的种子,对获得的M2、M3植株进行农艺性状观察和田间抗叶锈性鉴定。结果显示,不同浓度的EMS溶液处理种子共获得367个M1单株,处理浓度为1.0%时,接近半致死剂量,且M2表型突变频率最高,适宜突变筛选。在2 359个M2突变群体中,共筛选到表型突变体38个,表型突变频率1.61%;筛选出感叶锈病突变体53个,感病突变频率2.25%。在459个M3感病突变群体中,共鉴定出146个感病突变体,其中M36-2、M333-8、M333-9、M333-11、M344-4、M396-8这6个M3感病突变材料,遗传稳定率较高,达70%以上。为保证结果的可靠性,试验中还利用Lr19的分子标记对突变体进行分子辅助筛选。结果表明,EMS诱变是获得小麦感叶锈病突变体的有效手段,不仅为小麦抗叶锈基因的克隆和功能研究提供了重要的基础材料,还为小麦育种提供了新的种质。     

利用 F2 转 Bt 基因抗虫杂交棉新策略

通过不同抗虫棉品系间互交、后代选择的方法选育出聚合整合位点不同的Bt基因纯合抗虫棉种质,开展了培育以 F2 利用为目的的杂交种探索研究.类似 F2 世代 Bt 基因的不同分离类型,聚合有不同数目 Bt 基因的棉株在整个生育期对棉铃虫有显著的抗虫性.并且与单价抗虫棉时空表达一致,生育前期抗虫性好,毒蛋白表达量高,中、后期抗虫性有所下降,毒蛋白表达量降低.因此,用其作为亲本配置抗虫杂交种,是利用棉花F2 杂种优势的途径之一.     

A gene complex for annual habit in sugar beet (Beta vulgaris L.)

Annual habit in sugar beet has been shown to be controlled by a dominant gene, B, which induces bolting under long days without the cold requirement usually essential for biennial cultivars. The induction of bolting by B, however, is often influenced by both environmental and genetic factors. We studied the genetic basis for bolting suppression, caused by delayed planting, in lines derived from a cross between annual and biennial lines. The F₂ progeny of a late-bolting F₁ plant yielded an unexpected segregation ratio of annuality from monogenic inheritance when planted in late May, there being an excess of non-bolting biennials, although the expected segregation was observed in the late April sowing. Bolting suppression was caused by restricted daylengths due to delayed planting because the segregation was normal under artificially induced long days. The analysis with Idh1, an isozyme marker proximal to B, demonstrated that heterozygous B plants were more susceptible to bolting suppression due to delayed planting than homozygous B plants. The results suggest that bolting suppression was controlled by a number of genes responsible for long daylength requirement, one of which was closely linked to B and formed a gene complex for annuality. The annual habit was controlled singly by B under long days but modified by the genes for long daylength requirement under restricted daylengths. This revised version was published online in July 2006 with corrections to the Cover Date.     

甲基磺酸乙酯(EMS)处理对麻疯树(Jatropha Curcas L.)萌发种子的生理效应

分析了不同甲基磺酸乙酯(EMS)浓度(0、0.1%、O:5%、1.0%、1.5%和2.O%)和在浓度为1.2%时不同处理时间(O、8、12、16 h和20 h)对麻疯树萌发种子的生理效应.结果表明,EMS对麻疯树种子的半致死浓度为1.2%.当浓度达到2.0%时,种子受到严重伤害,成苗率为1.67%.用1.2%浓度诱变麻疯树种子,随着处理时间的延长,种子受到的伤害也随之加重,并且产生的矮化植株率也随之大幅度升高,在处理时间大于12 h时,得到的植株都是矮化植株,但当处理时间达到20 h时种子成苗率为0.由此看来,用EMS诱变麻疯树种子的合适浓度为1.2%,处理时间应选在8～12 h范围内.     

A marker-assisted analysis of bolting tendency in sugar beet (Beta vulgaris L.)

Bolting tendency in sugar beet varies among breeding lines and cultivars. Four crosses were made between breeding lines susceptible and resistant to bolting in order to study the genetic basis of bolting tendency. Bolting percentage in F₂, after 8 weeks of low temperature treatment, varied among the crosses, suggesting a complicated genetic control of bolting tendency. Different segregation ratios were observed, in particular, between families derived from the bolting F₁ plants and those from their non-bolting siblings, the former families showing a significantly higher bolting percentage than the latter. A marker-assisted analysis with seven isozyme loci, Ak1, Gdh2, Idh1, Lap, Mdh1, Pgi2 and Pgm1, revealed that a locus with marked effect on bolting tendency was located near Idh1. Because of a close linkage of Idh1 with B for annuality, the gene tagged by Idh1 appeared to be equivalent or similar to B′ for easy bolting allelic to B as reported by Owen et al. (1940). The results obtained suggest that the B locus may not only have an important role in determining growth habits but also control various degrees of bolting tendency in individual sugar beet plants. A linked pair, Ak1-Lap, and Pgi2 also were found to affect bolting tendency, although their effects varied depending on the crosses and families tested. This revised version was published online in July 2006 with corrections to the Cover Date.     

理化诱变小豆京农6号突变体的鉴定

选用小豆品种京农6号种子,分别采用甲基磺酸乙酯(EMS)(0.5%、0.9%和1.4%处理12h和24h)、电子束(100、300、600Gy)、60Co-γ(400Gy)诱变处理,将处理后的种子种于大田,鉴定后代植株性状的变异。观察表明,EMS诱变的变异类型最丰富、60Co-γ射线次之、电子束产生的变异类型较单一。EMS处理小豆以浓度0.5%和0.9%处理24h为宜;0.5%EMS处理的粒色和荚色变异突出,有鲜红、黄白、绿白粒色和黑荚、褐荚、黑褐荚变异;0.9%处理的叶形变异突出,有鸡爪叶、剑叶、肾形叶、小密叶等突变类型;电子束诱变后,M2变异率分别为4.09%、3.64%和2.22%。400Gy60Co-γ射线处理种子,后代变异率为7.23%。通过两年的鉴定筛选,获得937个EMS诱变M3代株系,934个60Co-γ射线和电子束诱变M2代株系,已得到株高、叶形、叶色、粒形、粒色、荚色、无分枝、多分枝、叶簇生、分枝簇生、光叶、蔓生、有限结荚习性、株型松散、育性、成熟特性等突变体材料1490份。本研究为小豆基因遗传分析、基因定位与克隆及其进一步的基因功能分析奠定了基础,为小豆育种提供了重要的材料。     

Transformation of tobacco with an Arabidopsis thaliana gene involved in trehalose biosynthesis increases tolerance to several abiotic stresses

Summary Trehalose (a non-reducing disaccharide) plays an important role in abiotic stress protection. It has been shown that using trehalose synthesis genes of bacterial origin, drought and salt tolerance could be achieved in several plants. A cassette harboring the AtTPS1 gene under the control of the CaMV35S promoter and the Bialaphos resistance gene was inserted in the binary plasmid vector pGreen0229 and used for Agrobacterium-mediated transformation of tobacco (Nicotiana tabacum). T0 plants obtained were analyzed by PCR for the presence of AtTPS1 gene. Thirty lines were positive and seeds were germinated on media with 6 mg/l PPT to obtain T1 plants that were grown in the greenhouse to obtain T2 seeds that were germinated on selective media. Lines which seeds showed a 100 % survival rate were considered homozygous transgenic T1 lines. Three lines were selected and gene expression confirmed by northern and western blots. Transgenic seeds were germinated on media with different concentrations of mannitol (0, 0.25, 0.5 and 0.75 M) and sodium chloride (0, 0.07, 0.14, 0.2, 0.27 and 0.34 M) to score their tolerance to osmotic stress. Assays were conducted to test the tolerance of transgenic plants to drought (measurement of water percentage as a consequence of water withdrawal), desiccation (measurement of water loss as a consequence leaf detaching) and temperature stresses (germination at 15 ^∘C and 35^∘C). Transgenic tobacco plant lines registered higher germination rates under osmotic and temperature stress situations than did wild-type plants. Responses to drought and desiccation stresses were similar for all plant lines. It can hence be suggested that the heterologous expression of TPS1 gene from Arabidopsis can be used successfully to increase abiotic stress tolerance in model plants and probably in other crops.     

EMS离体诱变和抗大花萱草叶枯病突变体的筛选

利用甲基磺酸乙酯(EMS)诱变大花萱草‘红运’愈伤组织和分化芽,目的是获得抗叶枯病愈伤组织和分化芽突变体。结果表明：0.50%~0.75% EMS处理愈伤组织60 min;0.75%~1.00% EMS处理分化芽60min,分别获得“半致死剂量”效应。对存活的愈伤组织进行毒素浓度递增的多步筛选(10% 30 d—20% 30 d);分化芽80%毒素21d的一步筛选。经定向筛选,初步获得抗叶枯病分化芽突变体103株,有待进一步进行抗病鉴定。愈伤组织未能获得抗病突变体再生植株。     

结球甘蓝自交系抽薹与开花性状配合力及遗传力分析

对6个甘蓝自交系的抽薹期与开花期性状配合力进行分析,并估算其遗传参数,为筛选时耐期抽薹品种提供理论依据.采用Griffing完全双列杂交方法Ⅳ对6个结球甘蓝优良自交系的抽薹期和开花期性状进行配合力和遗传力分析.结果表明:自交系05521-2和04M9-2的一般配合力均较好,具有作为耐抽薹亲本材料的潜力;抽薹期性状的广义遗传力和狭义遗传力分别为92.6%和50.04%;开花期性状的广义遗传力和狭义遗传力分别为92.09%和67.28%,抽薹期和开花期性状主要受加性效应基因控制,在早世代着手对结球甘蓝的抽薹期和开花期性状的选择是有效的.     

Early-bolting trait and RAPD markers in the specific monosomic addition line of radish carrying the e-chromosome of Brassica oleracea

The specific monosomic addition line of radish, Raphanus sativus, carrying the e chromosome of Brassica oleracea (2n = 19, e‐type MAL) with the genetic background of the late‐bolting cv.‘Tokinashi’ was produced by successive backcrossing of the original e‐type MAL of radish that showed early bolting in the genetic background of the cv. ‘Shogoin’. The early‐bolting trait specific to the e‐type MAL was constantly expressed in the backcrossed progenies (BC₂, BC₃ and BC₄), whereas the reverted radish‐like plants (2n =18) were gradually converted to bolting as late as ‘Tokinashi’. The added e‐chromosome expressed an epistatic effect against the genome of Japanese radish. Its early‐bolting trait was dominant to the late‐bolting trait of ‘Tokinashi’ which may be under the control of a few genes. Moreover, e‐type specific RAPD markers detected in eight primers were invariably transmitted in the backcrossed progenies by ‘Tokinashi’. From the analysis of the characteristics to the e‐type MAL and e‐type specific RAPD markers, it is suggested that the e‐added chromosome of kale (B. oleracea) was transmitted from generation to generation without any recombination with the radish chromosome. The gene(s) for the early‐bolting trait detected in this study may be useful for breeding work in radish, especially in the tropical areas.     

Inheritance of Bolting in Three Sugar Beet Crosses after Different Periods of Vernalization

The induction of bolting after different periods of vernalization was studied in the parental, F₁, F₂ and backcross generations (F₂× P₁ and F₂× P₂) of three crosses between five sugar beet lines. The parental lines represented different levels of bolting resistance. The populations were evaluated in replicated field or greenhouse trials. Generation mean analysis was used to quantify the genetic effects of bolting. The additive (d) parameter was predominant for those levels of cold treatment that stimulated the bolting to occur. The dominance (h) parameter was also shown to be important in most cases. In one cross, the additive × additive (i) type of epistasis was significant. The chances of detecting genetic effects are increased by exposing the plants to vernalization conditions which maximize the difference between two parental lines of each cross. Hence, it would be sensible to exploit both additive and non-additive genetic effects in any selection programme.     

冬瓜EMS诱变条件优化及突变体筛选

为研究甲基磺酸乙酯(EMS)诱变技术在冬瓜种质资源创制中的效果和探索适宜的冬瓜种子诱变体系,采用EMS磷酸缓冲液对冬瓜种子进行诱变处理,研究了处理浓度和诱变处理时间对冬瓜种子萌发和生长的影响,并对冬瓜M1、M2代突变体的田间生物学性状进行调查分析。结果表明,最佳的诱变条件为1.2% EMS诱变12 h,在此条件下,冬瓜种子发芽率、成苗率分别为54.67%、51.83%,接近半数致死浓度;在该诱变条件下大规模处理冬瓜种子10000 粒,催芽播种4752 粒,成苗4500 株,植株具有典型的EMS处理后的真叶畸形、黄化等特征,证明EMS诱导成功,自交授粉获得586 个自交瓜。种植M2代株系后获得叶、花、果实和株型等相关的突变体,突变率分别为27.22%、26.80%、20.47%、16.06%,冬瓜总突变频率为11.53%。本研究建立了冬瓜种子EMS诱变体系,构建了冬瓜突变体库,为冬瓜功能基因研究奠定了材料基础。     

EMS诱变谷子‘长农35号’M1代成熟期株型突变体的鉴定与分析

为建立谷子突变体库,以便在谷子分子生物学研究中应用,以生产上主要推广种植的谷子品种‘长农35号’干种子为实验材料,采用0.8%和1.0%甲磺酸乙酯(EMS)进行了诱变处理,试验共收获株型相关M1代突变体材料282份,其中,0.8% EMS诱变‘长农35号’获得株型突变体材料100个单株,可分为10个组;1.0% EMS诱变‘长农35号’获得株型突变体材料182个单株,可分为17个组。M1代成熟期株型相关性状突变体分析结果表明：1.0% EMS处理‘长农35号’所获突变体,在株高、穗下节粗、穗下第一节间粗和茎节数4个性状与对照有显著差异,而0.8%处理与对照差异不显著,因此,对于‘长农35号’来讲,采用1.0% EMS进行诱变处理更有利于多类型大量突变单株的获得。     

亚洲棉EMS诱变条件优化与突变体筛选

【目的】明确适于亚洲棉诱变的甲基磺酸乙酯(EMS)最佳浓度和处理时间,创制优异亚洲棉突变体。【方法】用体积分数0.4%~1.5%EMS溶液浸泡处理亚洲棉石系亚1号种子4~8 h,分析不同时间、不同浓度的EMS溶液处理对石系亚1号种子萌发和生长的影响,并对M1、M2代突变株的形态学特征进行了初步鉴定。【结果】0.6%EMS溶液处理8 h,石系亚1号种子的发芽率、发芽指数分别为51%、18.84,诱变效果最佳。以该参数大规模诱变处理约23 000粒石系亚1号种子,共筛选获得M1代变异材料5559株,突变频率为48.37%;M2代变异材料825份,突变频率为14.17%,其中叶型、株型的突变频率最高,分别为8.0%,5.9%;M3代变异株系57个,变异性状遗传频率为31.30%,M3代新增突变频率为18.26%。【结论】本研究建立了亚洲棉种子EMS诱变体系,构建了亚洲棉石系亚1号M2突变群体,获得了36份可稳定遗传的突变体株系,为棉花功能基因组研究奠定了材料基础。     

EMS诱变剂处理对苦荞种子萌发及主要农艺性状的影响

为探究不同甲基磺酸乙酯(ethyl methane sulfonate,EMS)浓度对苦荞种子生长发育的影响,加快苦荞品种改良的进程。本研究以‘川荞1号’、‘川荞2号’和‘川荞3号’3个品种作为试验材料,使用EMS对苦荞种子进行诱变。分别设置不同浓度梯度的EMS(0.4%,0.8%,1.2%,1.6%)处理苦荞种子,相应设置空白对照组。每个处理进行5次重复,室内调查萌发指标以及移栽后观察记录诱变植株的田间相关性状。结果显示:EMS除了对苦荞茎粗有一定的促进以外,随EMS浓度的增加,苦荞各品种全生育期延长。EMS对苦荞发芽率、根长、芽长、种苗鲜重、田间成苗率、株高以及产量方面影响显著,随着处理浓度的增加,各个指标呈递减趋势。说明EMS对苦荞植株的生长发育有明显的抑制效果。结合苦荞种子萌发相关性状、田间生长发育以及产量等方面考虑,综合得出EMS诱导苦荞适宜浓度为0.8%~1.2%。本研究为苦荞EMS诱变提供诱变时间和适宜浓度的参考。     

转Xa23基因水稻的白叶枯病抗性及其遗传分析

在分离克隆抗白叶枯病基因Xa23研究中获得大量转基因水稻材料。为了系统研究转Xa23基因水稻的抗病稳定性和遗传模式, 本文通过逐株进行抗白叶枯病接种鉴定、PCR和Southern blot分子检测, 对一批转Xa23基因水稻植株进行了T₀代到T₂代的跟踪分析。结果表明, Xa23基因的整合和表达, 使感病受体品种牡丹江8号获得抗病性。由于Xa23基因插入受体基因组的位点不同, 同是单拷贝插入的转基因T₀代抗病植株, 其抗病程度有明显差异。T₀代植株的抗病程度, 可以准确、稳定地遗传到T₁代和T₂代。单拷贝转基因植株分离群体的抗感植株分离比接近3∶1, 表明转Xa23基因遵循孟德尔单基因遗传模式。已获得2个纯合的单拷贝转基因抗病株系, 它们的抗病程度稍有差别, 将用于外源基因插入位置效应分析和杂交稻抗病育种。     

农杆菌介导的辽宁碱蓬胆碱单氧化酶基因CMO转化烟草提高抗氧化胁迫的研究

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