Induced chlorophyll mutations. I. Mutagenic effectiveness and efficiency of EMS, HZ and SA in mungbean

A systematic and comparative study on the frequency and spectrum of chlorophyll mutations induced by ethylmethane sulphonate (EMS) — an alkylating agent, hydrazine hydrate (HZ) — a base analogue and sodium azide (SA) — a respiratory inhibitor, was carried out in two mungbean varieties, namely, PDM-11 and NM-1. Awide spectrum of chlorophyll mutants was obtained in the M₂ generation. All these chlorophyll-deficient mutants were lethal except maculata, viridis and virescent. Chlorina followed by xantha types were predominant in both the varieties. EMS treatments induced the highest frequency of chlorophyll mutations followed by HZ and SA. The frequency of chlorophyll mutations was dose-dependent and increased with the mutagen concentration. Based on effectiveness in both varieties, the order of mutagens was HZ > SA > EMS. Two criteria viz., pollen sterility (Mp/S) and seedling injury (Mp/I) were taken into consideration to determine the efficiency of the mutagens. EMS was found to be the most efficient mutagen followed by HZ and SA. Moderate concentrations of the mutagens were the most effective and efficient in inducing mutations.     

草莓离体诱变及耐盐筛选技术初探

以‘全明星’草莓组培苗为试材,分别用甲基磺酸乙酯(EMS)、60Co-γ射线以及60Co-γ射线+EMS的复合诱变剂对叶片进行处理,并对‘全明星’草莓进行初步耐盐筛选。结果表明:EMS溶液处理草莓离体叶片适宜的浓度和时间是0.2%+2.0 h及0.4%+1.0 h,60Co-γ射线诱变处理的剂量率为30 R/min时,适宜的剂量为3~5 kR,复合处理的适宜剂量为60Co-γ(3 kR)+0.1%EMS处理2.0 h及60Co-γ(3 kR)+0.2%的EMS处理1.0 h。对3种处理间进行比较发现,各处理的叶片存活率和再生率都比对照有所下降。其中,复合处理对草莓叶片的生理损伤作用显著提高,叶片的再生率比单一诱变剂处理下降了10%~15%。再生植株经过初步NaCl筛选后,复合诱变处理再生植株的生物产量显著高于其它处理,盐害率和盐害指数也都显著低于其它处理,表现出较强的耐盐性,而单一诱变剂处理之间的差异不显著。     

Cytological effects of different mutagens in soybean [Glycine max (L.) Merrill]

The effect of gamma rays, ethyl methane sulphonate, and their three combinations on various cytological parameters in M1 generation was studied in two soybean cultivars, Pusa-16 and PK-1042. Combined treatments exhibited high meiotic abnormalities in Pusa-16 as compared with the individual treatments of ethyl methane sulphonate and gamma rays. In PK-1042 the highest frequency of meiotic abnormalities was exhibited by gamma rays, while in Pusa-16 the frequencies of univalents were high as compared with other meiotic abnormalities. Fragments/bridges, laggards, ring and rod bivalents were other types of meiotic abnormalities that were noticed in both the cultivars. The frequencies of irregular cells in the two cultivars were high in combined treatments followed by ethyl methane sulphonate and gamma rays. The highest percentage of pollen sterility was exhibited by the combined treatments and the lowest was exhibited by gamma rays. In Pusa-16, the increase in doses of gamma rays and ethyl methane sulphonate increased the pollen sterility percentage, while in the combined treatments, the increase was noticed up to the intermediate dose and then gradually decreased at higher doses. On the other hand, in PK-1042 the increase in ethyl methane sulphonate dose increased sterility percentage, while no definite trend was noticed in gamma rays and combined treatments.     

高能混合粒子场辐照小麦的突变效应分析

 【目的】分析高能混合粒子场（CR）辐照冬小麦所产生的突变效应,并与60Co-γ射线处理相比较,研究其诱变效率。【方法】以0、145、195、284和560 Gy剂量的CR和γ射线分别处理两个冬小麦品种ZY9和ZH7,室内发芽鉴定处理当代的损伤效应,田间筛选表型变异突变体并做考种分析。【结果】高能混合粒子场与γ射线处理对这两个小麦品种的生长抑制均存在明显的剂量效应,并且高能混合粒子场处理的损伤效应明显高于γ射线,ZH7的辐射敏感性大于ZY9,处理小麦的适宜剂量在200～300 Gy之间。M2代群体中出现了包括株高、穗型、生育期等多种性状变异,在适宜或稍高剂量时可以引起明显高于γ射线处理的有益突变。CR诱变ZY9的总突变频率和有益突变频率分别为5.42%～11.68%和2.75%～7.29%,ZH7则分别达到10.35%～22.12%和6.62%～10.49%,均高于γ射线处理频率。【结论】高能混合粒子场处理小麦能够产生比γ射线更高的相对生物学效应,而且辐射敏感性较高的品种,经处理后其后代诱发有益突变的频率较高,获得优良变异的几率也相对较高。     

EMS处理对枇杷试管苗表型及遗传物质的影响

以大五星枇杷幼胚培养获得的单一试管苗无性系茎尖为材料,用EMS进行化学诱变.结果表明:0.1％～0.9％EMS处理0.5～1.5h,试管苗死亡率很低;0.1％EMS处理0.5 h,植株明显高于对照,叶绿素含量也明显高于对照,为4.21 mg/g;经RAPD检测,诱变处理后的材料扩增谱带与对照间存在明显差异.     

几种化学诱变剂对玉米自交系的诱变效应

采用EMS,NaN3和苯甲酰胺复合剂、PYM3种诱变剂处理2个玉米自交系。结果表明：EMS,NaN3和苯甲酰胺复合剂在M1代引起了较大的生理损伤,并出现了较高频率的染色体畸变,PYM作为一种新兴的诱变刑对玉米的诱变效果并不显著,2个自交系品种对诱变剂的敏感性有差异。     

不同诱变处理对紫花苜蓿生理生化指标的影响

为了比较研究不同诱变处理方式对紫花苜蓿诱变效果的影响。试验通过60Co-γ射线、紫外线、甲基磺酸乙酯(EMS) 3 种诱变处理对供试4 种紫花苜蓿干种子进行诱变,以当代材料为研究对象,探讨诱变处理对苜蓿生理生化指标的影响。诱变处理后,3 种抗氧化酶活性以增加趋势为主。其中对SOD的影响,60Co-γ表现为促进作用,紫外线处理降低了SOD活性,EMS处理表现为低促髙抑作用;3 种诱变处理对POD和CAT的影响为60Co-γ辐射随处理剂量增加酶活性下降,紫外线处理则随着剂量增加酶活性增加,EMS处理则因品种不同而不同。诱变处理对苜蓿可溶性蛋白、可溶性糖和游离脯氨酸含量的影响随剂量及品种变化而不同,对MDA含量影响不显著。不同诱变处理显著提高了苜蓿叶片抗氧化酶活性,其中150 Gy 60Co-γ、90 min紫外线和0.4% EMS处理对苜蓿生理活性具有明显提高作用。     

玉米EMS诱变后代变化趋势的研究

[目的]为了研究玉米EMS诱变后代的变化趋势,以期得到有益变异和创造新的玉米种质。[方法]选取5种玉米自交系(340、2345、K12、136、9010)为供试诱变亲本,以EMS-石蜡油溶液为诱变剂,直接对花粉诱变得到诱变后代,研究M1、M2代的农艺性状与M3代种子的品质性状,分析诱变后代的变化趋势。[结果]各品种畸变率随EMS浓度增加而变大。M1代的出苗率和成株率明显低于对照,M2代百粒重明显低于对照,出苗率和成株率比M1代高,但比对照低。EMS对M3代种子营养含量的影响是非定向的,对各品种的影响不同。[结论]用EMS诱变玉米花粉效果显著,对于诱变后代的生理损伤和遗传性状的改变很大,但是不同材料对于EMS的敏感程度不同,故对于每种材料应选择合适的EMS浓度。     

浸种温度与化学诱变剂对芝麻种子发芽的影响

为了探讨浸种温度及化学诱变剂对芝麻种子发芽的影响,在不同浸种温度下,研究了甲基磺酸乙酯(EMS)和叠氮化钠(NaN3)不同(质量)浓度处理不同时间对芝麻种子发芽势、发芽率、发芽指数、活力指数的影响。结果表明,4℃浸种后,EMS处理时间超过4h或者EMS质量浓度大于10g/L时,3个芝麻品种均不能正常发芽,而25℃浸种后,10g/L EMS处理6h或15g/L EMS处理4h,3个品种仍能正常发芽。4℃和25℃浸种后,不同浓度NaN3处理下,3个品种均能正常发芽,但当6mmol/L NaN3处理12h时,4℃下浸种,3个品种的发芽率均低于50.0%,而25℃下浸种,3个品种的发芽率最低为69.3%。相同(质量)浓度、相同处理时间下,4℃浸种能更好地发挥EMS和NaN3的诱变作用。不同芝麻品种对EMS敏感性存在较大差异,对NaN3的敏感性差异较小。从诱变育种创造有益变异角度,建议4℃浸种24h后,以5g/L EMS诱变处理8h或4mmol/L NaN3诱变处理12h。     

Induced morphological mutants in soybean [<Emphasis Type="Italic">Glycine max</Emphasis> (L.) Merrill]

The seeds of two soybean cultivars viz., Pusa-16 and PK-1042 were treated with 15, 30, and 45 kR of gamma rays, three concentrations of ethyl methane sulphonate (EMS) (0.1%, 0.2%, and 0.3%), and their combinations. Various morphological mutants were recorded in M₂ and M₃ generations. These morphological mutants were named on the basis of the part of the plant body affected. Eight different morphological mutants, i.e., tall, dwarf, gigas, black pod, smooth pod, black spotted seed, bold seeded, and early maturing were isolated. Among them, early maturing and bold seeded mutants had the potential to be incorporated into breeding programs.     

EMS诱变小麦愈伤组织选择抗旱突变体的研究

以抗旱性差的小麦品种温麦6号和周麦17的花药愈伤组织和幼胚愈伤组织为材料,进行 EMS诱变处理,结果表明:EMS溶液处理花药愈伤组织相对分化率近50％的浓度和时间组合为 0．20％ (2-6h);处理幼胚愈伤组织相对分化率近50％的浓度和时间组合为(0．20％-0．40％) (2-4h)。 将EMS诱变处理后分化的224株再生植株接种到PEG浓度为80g/L的生根培养基中,进行抗旱筛 选,共得到13株抗旱植株,平均变异率为5．8％。花药愈伤组织和幼胚愈伤组织EMS溶液诱变处理的 适宜浓度和时间组合为0．20％ 4h。     

辣椒突变体库的构建及突变群体表型变异分析

选用辣椒自交系‘6421’作为构建突变体库的试验材料,分析甲基磺酸乙酯(EMS)处理对辣椒种子萌发和幼苗生理特性的影响,确定适宜的EMS诱变浓度,在此基础上构建辣椒突变体库,并对突变体库中突变群体的表型变异进行分析。结果表明:EMS质量分数为1.0%时,辣椒种子发芽率和幼苗成活率分别为41.01%和49.32%,接近半致死剂量,且该浓度下辣椒幼苗中超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)活性均达最大值,分别为246.45 U/g、33.99 U/g和158.10 U/(g·min),丙二醛(MDA)含量为对照的1.85倍;初步构建了含有562个家系辣椒突变体库,突变类型可归纳为叶、茎秆、果实、育性、生育期等5个类别,突变植株数分别占总突变植株数的12.73%、21.56%、6.92%、45.33%和13.46%。     

大豆EMS化学诱变处理条件分析

[目的]为使EMS(甲基磺酸乙酯)诱变效果最佳,获得高质量的诱变材料,其诱变时间和诱变浓度等基本条件分析是诱变处理的前提。[方法]采用化学诱变剂EMS处理大豆品种长豆18和黑豆品种长豆006,分析获得不同大豆品种的适宜诱变条件。试验设置2个品种、3个EMS浓度(0.2%、0.5%、0.8%)和3个诱导时间(4、8和12 h)共18个处理,以致死量达到50%为标准。[结果]大豆品种长豆18的适宜诱变条件以浸种4 h,处理浓度0.5%EMS,诱导时间8 h和浸种4 h,处理浓度0.5%EMS,诱导时间12 h处理最优;黑豆品种长豆006的适宜诱变条件为浸种4 h,处理浓度0.5%EMS,诱导时间8 h。[结论]大豆EMS化学诱变可适当提高致死剂量浓度作为大田处理浓度。     

EMS诱变六倍体小麦偃展4110的形态突变体鉴定与分析

 【目的】构建小麦EMS突变体库,为小麦功能基因组学研究准备基础材料。【方法】利用化学诱变剂甲基磺酸乙酯（ethyl methane sulfonate,EMS）诱变处理小麦品种偃展4110种子,将获得的M2代材料进行生物学性状与农艺性状鉴定,部分M3材料播种家系进行验证。【结果】对M2代全生育期田间表型进行观察鉴定,突变群体的表型变异率约为6.6%;获得了幼苗、叶、茎、穗及成熟期等生物学特性与主要农艺性状的变异体和突变体,变异类型丰富,特别是发现了自然突变中少见的变异类型,如株高在10-15 cm左右的特矮变异类型。【结论】本研究所构建的两个偃展4110 EMS突变群体较为理想,可望有效地被用于小麦功能基因组研究和小麦遗传改良中。     

用甲基磺酸乙酯(EMS)诱变的大麦浙农大3号突变体的筛选和鉴定

利用化学诱变剂甲基磺酸乙酯(ethyl methane sulfonate,EMS)诱变处理大麦品种浙农大3号的种子,然后种于大田,将获得的M2代材料进行生物学性状与农艺性状鉴定,入选的M3代材料继续进行验证.结果表明,实验获得了幼苗、生育期、叶、茎秆、穗及育性等生物学特性与主要农艺性状的突变体,变异类型丰富,突变群体的表型变异率约为7.46%.本研究构建的突变群体为大麦功能基因组研究奠定了基础,同时为大麦遗传改良提供了重要的材料.     

ISSR Analysis of Chlorophytum Treated by Three Kinds of Chemical Mutagen

Chlorophytum leaves were treated with three kinds of chemical mutagen, EMS, DES and NaN3 with different concentration to obtain the variation materials with excellent properties. The results showed that the genetic similarity coefficient of variation plants with EMS treatment was between 0.648 and 0.868, with NaN3 treatment was between 0.598 and 0.859, and with DES treatment was between 0.668 and 0.904, of which the mutagenic effects with 0.8% EMS, 250 mg. L-1 NaN3 and 0.3% DES on chloro- phytum were the best. ISSR molecular marker technique was used to analyze their genetic diversities. Total 392 polymorphic bands were obtained through 18 ISSR primers. Polymorphic ratio was 72.4%, which showed that DNA mutation took place in various degrees     

EMS处理对牛皮杜鹃组培苗叶片再生及耐热性的影响

该试验以长白山野生牛皮杜鹃组培苗为材料,探讨了EMS处理对牛皮杜鹃耐热性的影响和可能的耐热机理,为牛皮杜鹃推广应用及种质资源的创新奠定基础.结果表明,EMS浓度为0.1％处理2d时,叶片存活率48.92％.接近半致死剂量,可以将其作为耐热性突变体的筛选条件.随着温度的升高,诱变组植株叶绿素、MDA、Pro、SOD、POD活性、可溶性糖、MDA含量与对照均存在显著差异;相同温度时,诱变组植株叶绿素含量、可溶性糖含量、Pro,SOD和POD活性显著高于对照组.MDA含量显著低于对照组.EMS处理在一定程度上提高了牛皮杜鹃组培苗的的耐热性.     

不同色泽无花果果皮色素变化特征及其与果实糖分累积的关系

以5个不同果皮颜色的普通型无花果品种的秋果为试材,研究果实发育过程中果皮色素的变化,以及果皮色素含量与果实糖分累积的关系。结果表明:无花果果皮中叶绿素含量先升后降,类胡萝卜素含量先降后升,花色苷含量在成熟期呈持续上升的趋势。无花果果皮颜色是叶绿素、类胡萝卜素、花色苷等共同作用的结果,果实成熟期不同色素的含量因品种而异,红色和紫色品种果皮中的花色苷含量远高于黄色和黄绿色品种。紫色品种M121成熟期不仅花色苷和类胡萝卜素含量最高,而且叶绿素含量在5个品种中名列第二,一方面揭示了决定无花果色泽的色素多样性,另一方面表明了M121品种在抗氧化活性等方面具有的潜在的优势。无花果成熟过程中果实中的葡萄糖、果糖和蔗糖含量持续上升,可溶性糖的大量积累早于花色苷的大量积累。各品种叶绿素含量与可溶性糖含量均呈负相关,类胡萝卜素和花色苷与可溶性糖含量呈正相关。     

大麦TILLING体系在抗病基因研究中的应用

基因组靶向定位诱导损伤技术(Targeting Induced Local Lesions In Genomes,TILLING)是在化学诱变和PCR定向筛选基础上发展起来的检测点突变的反向遗传学研究方法,在多种重要农作物上都有应用。本研究使用化学诱变剂甲基磺酸乙酯(Ethyl methane sulfonate,EMS)处理大麦品种‘Tamalpais’,获得了2154个M2株系,同时开发了基于芹菜内切酶CEL I(celery juice extract)的酶切筛选体系。针对植物水杨酸抗病途径相关的两个重要基因EDR1和NPR1,检测到5个M2突变株系。序列分析表明,2个突变发生在内含子、1个NPR1基因的同义突变、2个EDR1基因的错义突变(His351Tyr和Pro556Ser)。本项研究为麦类作物反向遗传学研究奠定了基础。     

镉对两蕹菜品种生长的影响

[目的]探讨镉处理对不同品种蕹菜的影响差异。[方法]测量镉处理后蕹菜品种QLQ和T308株高、根长及叶绿素含量的变化。[结果]与对照相比,镉处理后2个品种株高均在0.05水平显著降低;QLQ根长在0.05水平显著增加,T308根长在0.05水平显著降低;QLQ中的叶绿素a、叶绿素b和总叶绿素含量在0.05水平显著降低,T308中只有叶绿素b含量在0.05水平显著降低。[结论]两蕹菜品种地上部对镉处理的响应相似,而根对镉处理的响应相差较大。