氨磺灵和秋水仙素诱导橡胶草四倍体效果的比较研究

为探讨不同诱变剂对橡胶草四倍体的诱导效果,以橡胶草幼芽为材料,采用浸芽法,比较分析氨磺灵和秋水仙素2种诱变剂不同浓度和处理时间对橡胶草多倍体的诱导效果及多倍体橡胶草的形态特征变化。结果表明：氨磺灵处理橡胶草的成苗率(60%~75%)和最高诱变率(14.08%)均高于秋水仙素处理（0~59%和5.56%）,其嵌合体率(27.27%)低于秋水仙素处理(75.00%),其中0.04 mmol/L氨磺灵处理 30 min,成苗率为71.00%,变异率14.08%,为最佳诱变条件。可见,氨磺灵比秋水仙素更适合用于橡胶草多倍体诱导。诱导后获得的四倍体橡胶草叶片厚度、气孔大小、花和种子大小均显著大于野生型二倍体植株,气孔密度则显著低于野生型植株,这些形态特征可辅助多倍体橡胶草快速筛选和鉴定。本研究初步探明了2种诱变剂对橡胶草染色体加倍的诱导效果,为今后橡胶草多倍体育种提供理论依据和技术支持。     

除草剂APM对小麦野二燕1号染色体结构和蛋白质组分的影响

利用农业上广泛使用的酰胺类除草剂甲基胺草磷,在1~4μmol/L浓度下,对野二燕1号小麦根尖分生组织细胞染色体结构变异及蛋白质组分变化进行了研究,结果发现,随着浓度升高,染色体结构变异率呈上升趋势。在3~4μmol/L浓度下处理的根尖分生组织细胞中,多极分裂细胞的出现频率为0.08%;染色体桥、断片和滞后染色体的出现频率为0.434%。双向电泳分析结果表明,约有10个蛋白质斑点产生变化,2个蛋白质斑点(8 kDa/PI 6.5,10kDa/PI 7.7)消失。染色体结构变异和蛋白质组分变化可以认为与甲基胺草磷的处理有关。     

秋水仙素诱导蚕豆及玉米根尖细胞染色体变异的研究

为进一步了解秋水仙素对农作物根尖细胞染色体畸变的遗传毒害效应,以地方品种敏感型绿皮小粒蚕豆和饲用玉米为材料,研究不同浓度秋水仙素（0、0.01%、0.05%、0.10%、0.15%、0.20%）及不同培养时间（24、48、72h）对2种作物染色体的畸变影响。结果表明：秋水仙素能诱发根尖膨大,当秋水仙素浓度为0.20%处理24h,蚕豆根尖膨大率100%;秋水仙素浓度为0.20%处理72h,玉米根尖膨大率65.8%;适当浓度秋水仙素（0.01%、0.05%）可促进蚕豆和玉米根尖细胞的有丝分裂,当浓度≥0.10%,反而抑制蚕豆和玉米细胞分裂。在染色体畸变类型中,微核最多,其次是染色体断片,最少是染色体桥。秋水仙素浓度0.20%,处理时间72h,蚕豆畸变率和玉米畸变率均最高,分别为11.92%和7.25%;秋水仙素浓度0.10%,处理时间72h,蚕豆染色体加倍指数最高为7.97%;秋水仙素浓度0.20%,处理时间24h,玉米细胞加倍指数最高为4.64%。     

啶磺草胺在小麦田施用时应严格控制用量

<正>小麦田常用除草剂啶磺草胺水分散粒剂(WG)作用靶标为乙酰乳酸合成酶(ALS)。石河子大学农学院通过研究新疆不同品种小麦的ALS对啶磺草胺WG的耐受性,确定啶磺草胺WG对不同品种小麦的安全浓度,筛选耐药性较好的小麦品种。科研人员利用室外盆栽法,将用清水浸泡24小时的小麦种子播种在花盆里,待幼苗长至3~4叶期后,喷施不同浓度的啶磺草胺WG,施药5天后提取     

滇丁香化学诱变的多倍体及其鉴定研究

为获得滇丁香新种质、选育抗性新品种,用不同浓度的氨磺灵溶液浸泡滇丁香种子,以种子下胚轴膨大视为变异株,统计变异率和发芽率,通过形态学观察、气孔观察、染色体计数及流式细胞仪检测确定其倍性。结果表明：15μmol/L氨磺灵浸泡种子24 h为滇丁香多倍体诱导最佳方法,变异率达60.1%,发芽率为66.7%,获得2株四倍体植株;25μmol/L氨磺灵诱导24 h,变异率为50%,获得1株八倍体植株。滇丁香变异植株在株高、叶长、叶厚方面均与对照植株呈显著差异,多倍体滇丁香植株株型矮小、叶片畸形较圆润且变厚、叶色较深、叶毛茎毛粗长。多倍体植株气孔的长和宽及保卫细胞的长和宽均显著大于二倍体植株,且气孔密度显著小于二倍体植株。经流式细胞仪检测,不同倍性植株DNA含量近似倍数关系。经染色体计数,滇丁香二倍体植株染色体数目为2n=44,四倍体植株4n=88。四倍体滇丁香丙二醛含量极显著低于二倍体,而脯氨酸含量极显著高于二倍体。     

青檀多倍体诱导试验初报

为获得具有优良性状的青檀四倍体植株,以青檀幼苗为试验材料,通过秋水仙素浸泡生长点的方法,研究了不同秋水仙素浓度和处理时间对青檀染色体加倍的诱导效果。结果表明：以0.4％—0.8％秋水仙素处理72ｈ诱导效果最佳,变异率最高达34.2％。经流式细胞仪和茎尖染色体鉴定,初次镜检青檀四倍体植株染色体数目为2n＝4x＝36,成功获得了青檀四倍体植株。     

由小麦×玉米获得的小麦DH系花粉母细胞减数分裂观察

研究了由小麦×玉米远缘杂交获得的小麦双单倍体DH2代花粉母细胞减数分裂过程。结果表明，大多数双单倍体小麦与异源六倍体普通小麦染色体数目相同，这是染色体经秋水仙素成功加倍的结果。同时在少数细胞中观察到了染色体的“块状移动”，在这些“块”中存在少数“染色体落后”现象，这可能是由于在染色体加倍过程中部分细胞染色体数目发生变异造成的。     

秋水仙素浓度、浸苗时间对烟草单倍体幼苗的加倍效应研究

为了提高烟草单倍体植株的染色体加倍率,采用二次饱和D-最优设计,以两片真叶期的烟草单倍体幼苗为材料,研究了秋水仙素浓度及其浸苗处理时间对染色体加倍的效应.结果表明,随着秋水仙素浓度的提高,加倍率呈开口向下的抛物线曲线变化趋势;加倍率与浸苗时间呈线性回归关系,加倍率随浸苗时间的延长而增加;0.346 9%的秋水仙素、浸苗72 h是烟草单倍体幼苗染色体加倍率最高(达54.18%)的最佳处理组合.     

NaCl胁迫对旱稻萌发及根尖细胞染色体行为的影响

旱稻是一种不耐盐碱、抗性较弱的作物。用不同浓度的NaCl处理旱稻种子,研究发芽情况和根尖细胞染色体行为。结果表明,随着NaCl浓度的升高,旱稻的发芽率、发芽指数、活力指数逐渐降低;NaCl浓度高于100 mmol/L时,引起根尖细胞染色体行为异常,异常类型有微核、多核、染色体落后、染色体粘连、染色体桥、染色体断裂等。且染色体异常行为与NaCl处理浓度呈正相关。     

苯甲酰胺对蚕豆根尖细胞变异性的影响

本文研究了苯甲酰胺对蚕豆根尖细胞微核细胞诱发率及染色体畸变率的影响。结果表明，苯甲酰胺可诱发蚕豆根尖细胞微核及染色体畸变的产生。在２０×１０＾６－２００×＾－６浓度范围内，随着浓度的增加，微核细胞率并未升高，反而有所下降，但在该浓度范围内，随着处理时间的延长，微核细胞率及染色体畸变色体畸变率明显上升。     

Efficient in vitro Chromosome Doubling During Beta vulgaris Ovule Culture

The effect of in vitro colchicine treatment of sugar beet ovules, after 7 days culture, on embryo formation, regeneration and ploidy of regenerated plants was studied with 5 concentrations of colchicine and 5 durations of treatment arranged as a 5 × 5 factorial in incomplete blocks. The best results were obtained with the shortest duration of treatment (5 hours) and the highest concentration of colchicine (0.4 %) giving 5.0 diploid plants per 100 ovules with 62.1 % of regenerated plants being diploid. Statistical analysis revealed that treatment effects could be separated into a toxic effect reducing embryo formation and a chromosome doubling effect affecting percentages of diploid regenerated plants. Toxic effects on embryo formation could be explained by simple exponential decay models, toxicity of the drug (decay constant) increasing linearly with duration of treatment. Duration of treatment had no effect on chromosome doubling percentages. The effects of colchicine concentration on chromosome doubling were explained by an exponential saturation model with spontaneous chromosome doubling of 8.1 % and saturation at 51.4 % diploid plants at 0.2 % and higher colchicine concentrations. In addition, treatments increased percentages of 4N and 6N plants from 0 % without colchicine to 10 % on average for treated ovules. A response surface model fitted to the total yield of diploid plants per ovule indicated that shorter durations of treatment and higher colchicine concentration may improve results.     

马铃薯高效染色体加倍方法建立与抗寒资源创制

In this work, different treatment combinations including colchicine concentrations and treatment methods as well as different time points was used to investigate the survival rate and chromosome doubling efficiency of potato tissue culture plantlets. The potato chromosome doubling by using colchicine had been successfully optimized. The potato plantlet stems treated with 0.1% colchicine for three days shaking at 120 r min^-1 showed the highest doubling efficiency due to its better contact to the colchicine solution. Compared with other potato chromosome doubling techniques, this method has much higher chromosome doubling rate, shorter time treatment and easier to operate, so that it could provide a higher efficient method for potato ploidy operation. In the meantime, compared with the diploid, the tetraploid interspecific hybrids showed differences in the morphological characteristic, which had higher plants, thicker stem, bigger petals and pollen grain. In addition, no significant difference was found between diploid and tetraploid interspecific hybrids in terms of cold resistance, but both significantly enhanced cold resistance compared with the common potato cultivar. Taken together, the doubled interspecific hybrids could sever for improving cold resistance of potato cultivars.     

Role of mitotic inhibitors and genotype on chromosome doubling of <Emphasis Type="Italic">Rosa</Emphasis>

Dinitroanilines represent a class of compounds that are widely used in herbicide formulations as they depolymerise plant microtubles, causing chromosome doubling. The potential of microtubule depolymerising herbicides trifluralin, oryzalin, and amiprophosmethyl (APM) for in vitro chromosome doubling of Rosa was studied. Five concentrations (0, 3, 6, 12 and 24 μM) and three exposure periods (12, 24 and 48 h) for each of the compounds were compared. Oryzalin, trifluralin and APM were not significantly different in their ability to induce chromosome doubling of R. hybrida cv Iceberg. At concentration of 6 μM and exposure period of 24 h, chromosome doubling of R. hybrida cv Iceberg was not significantly different with each of the polyplodising agents. At higher concentration (24 μM) and longer exposure period (48 h), 66.7% and 62.5% chromosome doubling was achieved with APM and trifluralin, respectively. However, the application of 6 μM oryzalin to R. persica (2n = 2x), R. hybrida cv Iceberg (2n = 3x) and R. hybrida cv Akito (2n = 4x), resulted in 60.0%, 6.3% and 0% chromosome doubling, respectively, which suggest that chromosome doubling is genotype dependent and plants with lower ploidy level have a higher propensity for chromosome doubling. Flow cytometry results at 18 and 24 weeks after herbicide treatment, indicated that the best time to test the treated plants was after 24 weeks.     

三氮苯类除草剂对水稻染色体结构、蛋白质含量及组分的影响

比较研究了不同浓度三氮苯类除草剂对水稻幼苗生长、叶绿素及可溶性蛋白质含量、根尖分生组织染色体结构、分生组织和叶绿体蛋白质组分的影响。结果表明,在0.1 mg /L Atrazine处理水稻后培养1周。幼苗高度由(10.8±1.7) cm（对照）降至 (8.2±0.7) cm,叶绿素含量由(1.07±0.013) mg/g FW（对照）降至(0.97±0.013) mg/g FW,     

Efficient Production of Doubled Haploid Plants through Chromosome Doubling of Isolated Microspores in Brassica napus

Three methods of chromosome doubling to produce doubled haploid plants from microspore cultures of Brassica napus were compared: colchicine treatment of microspore-derived plants, microspore-derived embryos, and isolated microspores. In the whole plant treatment, 53% of the treated plants set seed, but the treatment delayed plant growth and reduced seed set. When microspore-derived embryos were treated with colchicine, the doubling frequency was 32% (compared to 15% for spontaneous doubling). Direct colchicine treatment of isolated microspores resulted in a doubling efficiency of 70 % of the whole plants. This treatment also stimulated embryogenesis in microspore culture, leading to increased plant regeneration. Thus, direct chromosome doubling of isolated microspores is efficient and more than 10 000 doubled haploid plants have been produced in this manner in the past three years in order to accelerate the plant-breeding process.     

Antimicrotubule herbicides for in vitro chromosome doubling in Beta vulgaris L. ovule culture

In vitro chromosome doubling during ovule culture of sugar and fodder beets (Beta vulgaris L.) was studied with four anti-microtubule herbicides: amiprophos-methyl (APM), oryzalin, pronamide, and trifluralin at concentrations of 0–300 μM. Best chromosome doubling results were obtained by treatment of the ovules with 100 μM APM which produced 4.7 diploid plants per 100 ovules. Highest chromosome doubling was found with oryzalin using 1 μM, with trifluralin at 10 μM, and with pronamide at 10 μM producing 2.8, 2.0, and 2.0 diploid plants per 100 ovules, respectively. The APM treatments showed relatively low toxicity on embryo formation which in combination with a high chromosome doubling effect, resulted in up to 89 diploids per 100 plants regenerated. Oryzalin and trifluralin had more severe toxic effects, which reduced embryo formation, thereby lower percentages of chromosome doubled plants were obtained from these treatments. Pronamide had no significant toxic effect but it induced chromosome doubling at lower frequencies. Compared to colchicine, APM seems to be as efficient for chromosome doubling during beet ovule culture, but at molar concentrations 100 times lower than those used for chromosome doubling with colchicine. This revised version was published online in July 2006 with corrections to the Cover Date.     

The effect of colchicine on triticale anther-derived plants: Microspore pre-treatment and haploid-plant treatment using a hydroponic recovery system

In cereals, chromosome doubling of microspore-derived haploid plants is a critical step in producing doubled haploid plants. This investigation was undertaken to study the effect of incorporation of colchicine in the induction medium for anther culture, and the effect of colchicine on anther culture-derived plants of triticale grown under controlled greenhouse conditions. In the latter case, chromosome doubling of adult sterile plants derived from anther culture of fourteen triticale populations was attempted, where androgenetic plants with non-dehiscent anthers were cloned and subjected to the colchicine treatment, and then grown with the aid of hydroponics. The hydroponic system provided optimal conditions for recovery of the affected haploids from the toxic effects of colchicine treatment and all colchicine-treated plants survived. A topcross-F₁ (TC₁F₁) population with timopheevii cytoplasm produced the highest percentage of plants with seed-set either due to chromosome doubling by colchicine (98%) or spontaneous doubling of chromosome number (15%). Colchicine-treated anthers performed inferior than control in both induction and regeneration phases. One of the key observation of this study was the reversal from reproductive stage back to the vegetative stage which in turn enabled further cloning of haploid plants under hydroponic conditions once they were identified as sterile. The one hundred percent survival rate of in vitro-derived plants, 100% survival rate of colchicine treated haploid plants and the high chromosome doubling success rate (X = 82.3) observed in this study imply that a temperature-controlled greenhouse with an hydroponic system provides an efficient environment for inducing chromosome doubling of haploid plants in cereals. This revised version was published online in August 2006 with corrections to the Cover Date.     

Development of novel chromosome doubling strategies for wheat × maize system of wheat haploid production

Two chromosome doubling strategies were evaluated for producing wheat doubled haploids from wheat x maize crosses: (i) in vitro colchicine application to haploid embryos and (ii) colchicine treatment through postpollination tiller injections. In the in vitro approach the haploid embryos were rescued on medium containing colchicine (at concentrations of 0.2, 0.3, 0.4 and 0.5%) and moved to a colchicine‐free regeneration medium 48 h later. Embryos exposed to 0.5% colchicine had 91.67% of their regenerated plants showing chromosome doubling. In the tiller injection approach, different concentrations (0.5, 0.75 and 1.0%) of colchicine solution, which also contained 2,4‐D (100 ppm), were injected into the uppermost inter‐node of crossed tillers 48 and 72 h after pollination. The chromosome doubling efficiency varied from 33 to 100%, with 1% treatment being the most effective. No chimeras of doubled/haploid sectors were observed in the case of the tiller injection treatment and all the florets showed seed set in the doubled plants. Stomatal guard cell length provided rapid, early‐stage and unambiguous analysis of ploidy level on the basis of 10 guard cell observations per plant.     

RAPD analysis of sporting and chimerism in chrysanthemum

Summary The potential of colchicine and the microtubule depolymerizing herbicides trifluralin, oryzalin, and amiprophosmethyl (APM) for in vitro chromosome doubling during B. napus microspore culture was studied. Colchicine was administered during the first 6, 12 or 24 h of culture with 8 different concentrations up to 3 mM, and herbicides at 6 different concentrations up to 30 M for 12 h.Treatments with moderate concentrations of colchicine (3–100 M) produced a small increase in embryo production, while concentrations above 300 M were toxic. Colchicine treatment for 12 h resulted in higher embryo production than treatment for 6 and 24 h. Duration of treatment and concentration of colchicine both had a significant effect on the chromosome doubling. The highest diploidization rates (94% diploid regenerants) were seen after 24 h treatment with 1 mM colchicine.All three herbicides were similar to colchicine in terms of their effect on embryo formation and chromosome doubling comparable to the one of colchicine, but at concentrations approximately 100 times lower. APM was less toxic than trifluralin and oryzalin, but no significant difference in chromosome doubling efficiency was detected between the compounds. The 12 h treatment resulted in a maximum of approximately 65% diploid regenerants with all three herbicides, but APM may have an advantage because of its less toxic effects. Prolonged treatment with APM (20–24 h) may produce 95–100% diploid regenerants.Abbreviations APM amiprophos methyl - DMSO dimethyl sulfoxide