甲基磺酸乙酯（EMS）对萝卜早期发育性状的诱变效应

甲基磺酸乙酯(EMS)诱变育种有着诱变频率高、突变性状多及破坏性小等优点被广泛用于多种作物诱发突变育种。本研究利用不同浓度的EMS对不同品种的萝卜种子进行浸种处理后,发现EMS对萝卜的种子发芽率、萝卜苗根部长度及田间性状等指标上的诱变效应。研究结果表明随着EMS浓度(0.2%~2%)的增加,种子发芽率急剧降低且萝卜苗期根长明显变短,整体呈显著的抑制生长作用。不同品种间对EMS的敏感度依次顺序为短叶-13>夏抗40>春白11-58>双红一号。以半致死浓度为EMS浓度选择标准,确定了EMS处理不同品种萝卜间的适宜浓度为0.4%~0.6%。通过田间性状调查发现0.5%EMS处理短叶13萝卜种子下播大田后,相比对照处理品种表现子叶卷曲及撕裂、真叶增厚及黄化等情况,进一步表明EMS对短叶13萝卜存在的诱变效应。本研究确定了化学诱变剂EMS对萝卜诱变的半致死剂量,初步确认EMS对萝卜早期生长发育性状的诱变效应,将为后期的萝卜诱变育种研究提供研究材料和参考。     

Genetic relationships among three chlorophyll-deficient mutants in peanut,Arachis hypogaea L.

Summary The genetic relationships of three chlorophyll-deficient mutant peanuts, lutescens (lu), aureus (au), and virescent (v) were studied under field and greenhouse conditions. The F₁ plants from crosses between these mutants produced phenotypically normal green. In F₂, aureus X virescent segregated 675 normal green : 225 virescent : 45 aureus : 15 virescent aureus : 64 seedling lethal, and lutescens X virescent segregated 45 normal green : 15 virescent : 3 lutescens : 1 seedling lethal. (Lutescens peanuts were seedling lethal in the field.) As previously reported, the F₂ of aureus X lutescens gave 225 normal green : 15 aureus :15 lutescens : 1 seedling lethal. The three chlorophyll-deficient factors (au, lu, and v) show independent inheritance. The recessive combinations from the parental types between aureus and virescent and between aureus and lutescens would produce plants with a combination of their respective parental characteristics, but the recessive combination between lutescens and virescent was nearly albino. The v-au and lu-au seedlings have a longer life span than the v-lu seedling has. The genotypes for the three mutants are tentatively identified as lutescens VV Au ₁ Au ₁ Au ₂ Au ₂ lu ₁ lu ₁ lu ₂ lu ₂ L ₁ L ₁ L ₂ L ₂, aureus VV au₁au₁ au₂au₂ Lu₁Lu₁ Lu₂Lu₂ L₁L₁ l₂l₂, and virescent vv Au₁Au₁ Au₂Au₂ lu₁lu₁ Lu₂Lu₂ l₁l₁ L₂L₂.     

Variation in the physiological sensitivity of maize inbreds to EMS a possible correlation with alcohol dehydrogenase activity

Three inbred lines of maize, AD-1, AD-7 and AD-19, showed significant differences in their physiological sensitivity to ethyl methanesulfonate (EMS). Among the seedling parameters tested, the width of the leaves was the most sensitive and showed the best correlation with the results obtained at maturity. Plant weight was also a good seedling parameter. The physiological sensitivity to EMS was negatively correlated with alcohol dehydrogenase (ADH) activity in the scutellum of the three lines. A possible explanation for this correlation is discussed; however, it may have been due to chance.     

Modification of amino acid composition of endosperm proteins from in-vitro-selected high lysine mutants in rice

Summary Endosperm protein mutants in rice may be recovered by biochemical selections with inhibitory levels of lysine and threonine. Among the phenotypes recovered from in vitro selections are lines with increased protein and percent lysine in the protein. This work was designed to identify changes in proteins of rice mutants and to further our understanding of the mechanisms of lysine plus threonine selections in rice. Among the most obvious amino acid changes in mutants was a higher lysine level in all protein solubility fractions and a decrease in tyrosine. Methionine and glutamate are reduced in some protein fractions. However, methionine is significantly higher in the mutant than the control in the glutelin fraction. Several other aspartate pathway amino acids are higher in the mutant than the unselected controls. Separation of proteins in SDS-PAGE gels showed shifts in the protein profiles in the mutants, including a decrease in the major 30 kDa low lysine globulin component, and an increase in several high-molecular-weight components, approximately 60–100 kDa. Increases in the lysine content of proteins of different solubility classes and different proteins within classes are detailed.     

EMS诱变甘蓝型油菜M_2代群体的表型突变研究

利用4种浓度EMS处理甘蓝型油菜NJ7982种子,选取其中诱变效果好的处理(0.4%EMS),研究其后代突变型和突变频率。对4.8万株的M2代群体鉴定结果表明,6种器官性状在群体中均出现了突变。子叶突变性状包括3子叶、子叶黄化等,占群体的0.22%;叶片突变性状包括黄化叶、白化叶、紫色叶、上卷叶、下卷叶等,占0.74%;花器突变性状包括紫色花蕾、死蕾、3花瓣、6花瓣、白色瓣、花瓣黄白镶嵌、花瓣皱缩、完全不育、部分可育等,占9.38%;株型突变性状包括矮秆、紫茎等,占4.98%;角果突变性状包括粗角、长角、紫角等,占2.79%;种皮黄色,占0.40%;总的表型突变频率为18.51%。这些遗传多样性的突变材料,为甘蓝型油菜种质创新及品种遗传改良提供基础性材料。     

Effectiveness and efficiency of ethyl methanesulphonate and ethylene oxide for the induction of mutations in rice

A comparison of the mutagenic efficiency and effectiveness of ethyl methanesulphonate (EMS) and ethylene oxide (EO) on two different genotypes of rice showed that the mutagenic efficiency sharply decreased with increase in the concentration of EO whereas the efficiency of EMS increased with an increase in the concentration. The effectiveness was inversely proportional to dose of EO whereas it varied little with an increased dose of EMS.     

Fine characterization of OsPHO2 knockout mutants reveals its key role in Pi utilization in rice

Previous research using forward genetics approaches demonstrated that OsPHO2 regulates multiple phosphate-starvation responses in rice. In this work, we finely characterized two independent OsPHO2 knockout rice mutants under inorganic phosphate (Pi)-sufficient conditions. The ospho2 mutants exhibited defects in growth and reproductive development in the whole growing period. The cells in the elongation zone of ospho2 seedling roots were much shorter than those of the wild type. The phosphorus concentration in the blades of ospho2 mutants was 5.8-fold higher than those of wild-type plants, whereas it was only slightly higher in the sheaths, culms, spikelets, and seeds. Furthermore, Pi levels in the ospho2 mutants were highest in the oldest leaf and lowest in the youngest leaf, whereas there was no significant difference in the corresponding leaves of wild-type plants. These results suggest that ospho2 mutant phenotype results from a partial defect in Pi translocation and remobilization in the shoot of rice. This study thus provides evidence that OsPHO2, which functions at the downstream of OsPHF1, modulates Pi utilization by regulating the expression of Pht1 transporters in rice.     

裸燕麦EMS突变体库筛选与分析

燕麦是重要的粮饲兼用作物,构建燕麦EMS突变体库对燕麦功能基因组学研究和遗传改良有重要意义。本试验利用化学诱变剂甲基磺酸乙酯(EMS,ethyl methane sulfonate)处理燕麦品种花早2号,获得了4083株M1材料;对其中2000个单株种植了M2株行,进行全生育期调查,鉴定其表型变化;对2份黄化苗突变材料种植了M3家系,进行相关突变性状的稳定性验证。结果表明,燕麦经EMS处理后代变异巨大,在M2发现表型突变材料196份,变异率为9.8%,变异类型非常丰富,包括幼苗习性、叶片性状、分蘖、株高、穗部形态及成熟期等突变株系。M3证实突变的黄化苗特性可以稳定遗传。本研究建立了燕麦EMS诱变体系,获得的燕麦变异类型丰富,为燕麦功能基因组学研究和燕麦遗传改良奠定了材料基础。     

Physiology and proteome responses of two contrasting rice mutants and their wild type parent under salt stress conditions at the vegetative stage

Salinity is one of the major environmental limiting factors that affects growth and productivity of rice (Oryza sativa L.) worldwide. Rice is among the most sensitive crops to salinity, especially at early vegetative stages. In order to get a better understanding of molecular pathways affected in rice mutants showing contrasting responses to salinity, we exploited the power of 2-DE based proteomics to explore the proteome changes associated with salt stress response. Our physiological observations showed that standard evaluation system (SES) scores, Na⁺ and K⁺ concentrations in shoots and Na⁺/K⁺ ratio were significantly different in contrasting mutants under salt stress condition. Proteomics analysis showed that, out of 854 protein spots which were reproducibly detected, 67 protein spots showed significant responses to salt stress. The tandem mass spectrometry analysis of these significantly differentially accumulated proteins resulted in identification of 34 unique proteins. These proteins are involved in various molecular processes including defense to oxidative stresses, metabolisms, photosynthesis, protein synthesis and processing, signal transduction. Several of the identified proteins were emerged as key participants in salt stress tolerance. The possible implication of salt responsive proteins in plant adaptation to salt stress is discussed.     

Mutants lacking glutelin subunits in rice: mapping and combination of mutated glutelin genes

Nine mutant lines lacking glutelin subunits were selected from M₂ seeds of about 10000 M₁ plants mutagenized with gamma rays or EMS and from 1400 mutant lines selected originally for morphological characters. There were three types of mutants, one line lacking the largest subunit among four minor bands of glutelin acidic subunits (Type 1), five lines lacking the second largest subunit band (Type 2), and three lines lacking the third largest subunit band (Type 3). Mutants lacking the smallest subunit band were not found. Type 1 lacked 2 of the 10 spots of glutelin acidic subunits separated by two-dimensional electrophoresis and 1 of the 11 spots of the 57-kDa glutelin precursor. Type 2 lacked 2 spots of acidic subunits and 1 spot of the 57-kDa glutelin precursor, and had low amounts of 1 of the 8 spots of glutelin basic subunits. Type 3 mutants lacked each of 1 spot of the acidic subunits and glutelin precursor and had low amount of 1 spot of the basic subunits. Genetic analysis of the mutated genes showed that these mutant characters were controlled by single recessive genes named glu-1, glu-2, and glu-3, respectively. Mutated genes of different lines of the same type were found to be at the same locus. RFLP analysis of F₂ plants between the mutant lines and cv `Kasalath' indicated that glu-1 is on chromosome 2, glu-2 on chromosome 10, and glu-3 on chromosome 1. These mutant genes were combined by crossing, and a line lacking the 3 minor bands of the glutelin acidic subunits was developed. However, the total glutelin content of this line was not remarkably reduced, showing a only 13% decrease. Received: 1 April 1996 / Accepted: 14 June 1996     

Colchicine-like effect of diethylstilbestrol (DES) on mammalian cells in vitro

Diethylstilbestrol (DES), a synthetic estrogen, showed colchicine-like effects in vitro on cells of the cell lines such as Chinese hamster fibroblast of thymus origin (CHT), rat liver (DL), rat erythroblastic leukemia (EDEN-1/TC) and HeLa-S3. Metaphase arrest was induced 3 h after treatment with 15 microgram/ml of DES and polyploid or polynucleated cells were prominently observed more than 24 h after treatment. The arrest, however, was reversible when the agent was removed from the medium. Tetraploid karotypes induced by DES in CHT cells consisted of all double sets of diploid chromosome constitution except one chromosome marker. By clonal selection, several hypotetraploid sublines were successfully isolated from a CHT cell population after the treatment with 15 microgram/ml of DES for 48 h. Some comparative studies of cytological effects of DES with those induced by colcemid indicated that the DES effect was also a mitotic inhibition similar to colchicine.     

不同水分管理方式下水稻的水分利用效率与产量

采用温室微区试验研究常规水作、裸地旱作、覆膜旱作和覆草旱作等土壤水分管理下水稻生长及其对水分的利用状况。结果表明,几种旱作水稻的需水量为349-473mm,常规水和水稻需水量为762.5mm。旱作水稻的水分籽粒和干物质生产效率为0.899-1.273g·kg^-1和1.655-2.321g·kg^-1之间,而相同条件下常规水作水稻水分的籽粒和干物质生产效率为0.766g·kg^-1和1.459g·kg^-1左右。覆草旱作水稻可以获得相当于常规水作水稻90％的经济产量。     

The nature of reverse mutations in Drosophila melanogaster

A selection system for the screening of reversions has been constructed and used to test reversions of lethals located in the proximal region of the X chromosome of Drosophila and of Kpn mutations.Spontaneous and induced reversions have been screened, X-rays and ethyl methanesulphonate (EMS) being the mutagens used in the induction experiments.No genuine back-mutation was found in 6·10⁵ gametes scored. Sterile reversions of all four lethals tested were obtained. Their frequency suggested that at least in three of the lethals the sterile reversions represented “escapers” of the lethal effect rather than true revertants.Three fertile reversions of l^x4 were found and analyzed. All three were autosomal suppressors, located on the second chromosome, allelic to each other, dominant in males and recessive in females.One fertile reversion of l^3DES was found to be an X-linked suppressor. It is suggested that this suppressor is a Y-suppressed lethal, showing a V-type position effect, resulting from an aberration included in the proximal heterochromatin of the X chromosome.Reversions of Kpn were obtained at a similar rate to that found in previous reports²².The absence of true back-mutants in our experiments, in contrast to findings in previous reports, is discussed. From the existing literature on spontaneous and induced back-mutations in Drosophila melanogaster it appears that for several mutations the rates of forward and back-mutation are of the same order of magnitude. It is suggested that reported cases of back-mutations represent mainly inter- and intrachromosomal recombination in duplicated regions rather than mutational events and that the frequency of true back-mutation in Drosophila is usually of an order of magnitude, similar to that known for microorganisms and fungi.     

生境异质度对稻田捕食性天敌及水稻害虫的生态调节有效性

【目的】探索生境高异质度对稻田捕食性天敌及水稻害虫的生态调节有效性,了解这种策略是否会引起其他植食性昆虫成为水稻重要害虫的风险。【方法】2017-2018年连续2年种植单季稻,在稻田边缘种植花生与大豆,构建高异质性边缘生境稻田(rice paddy with high heterogeneous marginal habitats, HHR),调查HHR稻田与简单低异质性边缘生境稻田(rice paddy with low heterogeneous marginal habitats, LHR)中捕食性天敌与水稻害虫功能团的发生规律与相关性,计算益害比。【结果】2017年在HHR稻田中采集到捕食性天敌40种,1 667头;在LHR稻田中采集到捕食性天敌30种,991头。2018年在HHR稻田中采集到捕食性天敌33种,1 384头;在LHR稻田中采集到捕食性天敌34种,1 031头。HHR与LHR两类稻田中获得的捕食性天敌群落重要值P_i≥0.01的物种相似度很高,优势种相似。2017年HHR稻田的捕食性天敌物种丰富度明显高于LHR稻田,这种差异主要由群落重要值P_i<0.01的种类引起。2018年两类稻田的捕食性天敌物种丰富度没有差异。单位样方面积内的捕食性天敌个体数量,HHR中明显高于LHR,这种差异在2017年的分蘖期与成熟期达显著水平(P<0.05),在2018年的开花期极显著(P<0.01)。2017年在HHR中采集到水稻害虫22种,637头;在LHR中采集到水稻害虫19种,743头;物种相似性系数0.88。2018年在HHR中采集到水稻害虫16种,1 011头;在LHR中采集到水稻害虫16种,2 014头;物种相似性系数0.75;主要害虫物种组成结构相同。水稻害虫数量发生的时间动态分析表明,在分蘖期,虽然HHR稻田中的水稻害虫数量明显高于LHR(P<0.05),但此期害虫的数量不多,发生较轻。在孕穗期、开花期与成熟期,HHR稻田中的水稻害虫数量明显低于LHR稻田,这种差异在2017年的成熟期与2018年的孕穗期极显著(P<0.01),在2018年的成熟期差异显著(P<0.05)。2017年HHR和LHR中个体数量益害比N_pi值分别为2.62和1.33;2018年分别为1.37和0.51。【结论】具有高异质性边缘生境的稻田,能提高系统对捕食性天敌物种的涵养潜力,显著提高稻田捕食性天敌个体数量,提高益害比,具有更好的控制害虫的物质基础,促进捕食性天敌对水稻害虫的生态控制效能,不会引起其他植食性昆虫演变为水稻重要害虫风险,可为保护稻田生态系统天敌发挥生态效能提供可借鉴的策略与方法。     

1,2,4-三氯苯对水稻种子萌发及幼苗生长的毒性机理

以水稻为试材,采用水培方法研究了1,2,4-三氯苯(1,2,4-TCB)对水稻种子萌发、幼苗生长、叶绿素含量、蛋白质含量、脯氨酸含量、细胞膜通透性的影响.结果表明,水稻种子的萌发率和活力指数随1,2,4-TCB处理浓度的升高而逐渐降低,呈现一定的浓度-效应关系,高浓度(80和100 mg·L^-1) 1,2,4-TCB处理组水稻种子的活力指数仅为对照组的7. 89％和1.92％,水稻幼苗的株高和根长均受到抑制,表现出一定的浓度-效应和时间-效应关系.水稻叶片的叶绿素a、叶绿素b、总叶绿素和蛋白质含量均随1,2,4-TCB浓度的升高而降低.水稻叶片游离脯氨酸含量和细胞膜的通透性随处理浓度的升高而增加.当1,2,4-TCB处理浓度＞15 mg·L^-1时,游离脯氨酸含量和叶片细胞膜的通透性急剧增加,与对照组相比各组差异均显著(P＜0.01).这表明1,2,4-TCB对水稻幼苗的损伤是一个复杂的过程,其机理可能与1,2,4-TCB使植物细胞膜、细胞内蛋白质和细胞器结构和功能损伤有关.     

北方稻田生态系统水量平衡及水分效率研究

１９９３～１９９５年研究了５种不同模式水稻田生态系统水量平衡及水分效率结果表明,不同水稻田模式其总耗水量之间有明显差异,其中节水模式和节水节肥模式较常规模式节省灌溉水达１５～２３％,水分生产效率增加３０％以上．各模式蒸发蒸腾耗水量在同一生长季内基本相同．田间结构及调控管理对其无明显影响实测水稻生育期田间蒸发蒸腾量与计算的可能蒸发蒸腾量相差不过５％。     

Mutants having a low content of 16-kDa allergenic protein in rice (Oryza sativa L.)

Summary Rice mutants containing low levels of the 16-kDa allergenic protein, which is the main allergen in the rice grain for patients of atopic dermatitis due to the intake of rice, were screened, and 4 independent mutant lines with small amounts of this protein were found by SDS gel electrophoresis and immunoblot analysis. These mutants were grouped into two types. Two mutant lines, 85KG-4 and 86RG-18, contained low levels of the 16k-Da and 26-kDa polypeptides and a high level of the 57-kDa polypeptide. The 16-kDa polypeptide content of these mutants was about half that of the original cultivars. Homozygous lines were developed, and these showed normal growth and seed set. The other 2 mutant lines, 87KG-970 and 89WPKE-149, showed traces of the 16-kDa and 26 kDa polypeptides and contained a high level of the 13 kDa polypeptide. The homozygous plants of this type were sterile. All of the mutant lines had floury endosperms. Genetic analysis suggested that low 16-kDa polypeptide content is controlled by a single recessive gene. Attempts to separate of the genes for low 16-kDa polypeptide content and floury endosperm by crossing with the original cultivar were unsuccessful, suggesting the tight linkage of these two genes or pleiotropism of a single mutated gene. The relationship between low 16-kDa polypeptide content and the floury character and the possible use of the mutant as a low allergen rice are discussed.     

Internal efficiency, nutrient uptake, and the relation to field water resources in rainfed lowland rice of northeast Thailand

Rice-based (Oryza sativa L.) rainfed lowlands are the major cropping system in northeast Thailand. Average yields are low, which is generally explained by frequent drought events, low soil fertility, and poor fertilizer response. However, neither the relative importance of these factors nor their interaction is well understood. Therefore, we analyzed an existing database on fertilizer trials conducted between 1995 and 1997 at eight different sites in northeast Thailand with the objective to determine indigenous nutrient supplies, internal efficiencies, and recovery efficiencies of applied nutrients in rainfed lowland rice. Of particular interest was the effect of variety type (traditional) and water supply on these components. Comparison of N, P, and K concentrations in grain and straw (average N–P–K grain concentration of 11.0–2.7–3.4 g kg⁻¹; average N–P–K straw concentration of 5.2–0.9–16.4 g kg⁻¹) in the traditional-type varieties used at all trial sites with literature values showed no differences for these parameters between traditional and modern-type varieties or between irrigated and rainfed environments. In contrast, internal efficiencies of N, P, and K (average IEN: 46 kg grain per kg N uptake; IEP: 218 kg grain per kg P uptake; IEK: 25 kg grain per kg K uptake) were much lower than reported for irrigated systems, and the difference was greatest for K, which is mainly accumulated in the straw. Indigenous nutrient supply (average INS: 38 kg ha⁻¹; IPS: 10 kg ha⁻¹; IKS: 89 kg ha⁻¹) and recovery efficiency (average REN: 0.28 kg kg⁻¹; REP: 0.13 kg kg⁻¹; REK: 0.49 kg kg⁻¹) were low but comparable to the lower values reported from irrigated systems. Average seasonal field water resources seemed to reduce the indigenous nutrient supply but had no or little effect on internal efficiency and recovery efficiency. We concluded that the main reason for the low system productivity without and with fertilizer in northeast Thailand is the dominant use of traditional-type varieties with low harvest indices, which was the dominant cause for the observed low internal nutrient efficiency. Therefore, intensification of rainfed systems through substantially increased nutrient inputs can be recommended only where varieties with an average harvest index of close to 0.4 or higher are available.