植物诱变技术的研究进展

植物诱变技术是指利用外界因素加快物种遗传变异,在短期内获得有利用价值的突变体,为培育新种质、新品种及基因功能的研究等创造条件。相对于自然的选择,诱变技术具有高频率、广突变、周期短等特性。主要论述了常用的化学诱变、物理诱变、空间诱变和生物诱变等诱变手段,并详细介绍了上述诱变技术的诱变机理、生物学效应、常用的诱变方式及其在植物应用中的研究现状。针对现行各种诱变技术的不足提出了今后努力的方向。     

康乐霉素C产生菌的微波诱变

以物理因子微波对免疫抑制剂康乐霉不C的产生菌进行了菌种诱变,研究了微波处理的条件、照射的时间及剂量,获得康乐霉素C生产能力高于出发菌株180%的M-13-21。     

微波对阿维拉霉素产生菌诱变效应的研究

阿维拉霉素产生菌SV微波诱变效应的研究发现：菌株SV对微波敏感（微波处理60s,SV菌株存活率低于10％）,菌落形态变化大。微波诱变处理最佳作用方式为培养皿不加盖、快速冰上冷却,最佳处理时间为50s（其正突变率25．3％）。通过微波诱变处理、阿维拉霉素推理筛选,最终获得一株稳定性良好,阿维拉霉素产量达到21．5mg／L,较出发菌株提高119．4％的突变株SV-15。     

激光诱变育种的研究概况

阐述了激光诱变育种机理的研究进展,激光诱变在农作物、畜牧兽医、工业微生物育种上的应用及已取得的成果,并介绍了激光诱变育种的一般操作过程和激光诱变育种的研究前景。     

离子束诱变育种研究及应用进展

阐述了离子束诱变的生物效应,研究中有待解决的问题,介绍了离子束诱变在农业、工业育种上的研究进展及成果,并展望了今后的研究前景。     

一株感染家蚕的高致病性白僵菌的筛选及复合诱变

【背景】白僵蚕中的生物活性物质在医疗、保健品及化妆品行业有着广泛的应用。目前,许多人工养殖僵蚕基地在实际生产中使用的菌种多为未进行纯化优选的自然感病死亡僵蚕孢子粉且无固定的施用浓度,使得蚕的僵化死亡率难以保证。提高白僵菌菌株的致病力并筛选性状优良的高毒力菌株是工厂化生产白僵蚕研发的重要方向。【目的】利用紫外-微波复合诱变技术筛选高毒力菌株,为僵蚕工厂化生产提供优良菌株。【方法】利用孢子稀释法从山西省养殖农户中自然感染白僵菌的家蚕中分离获得一株原始白僵菌,运用紫外-微波复合的方式对该菌株进行诱变,并比较诱变前后菌株的产孢量及对家蚕的致病力。【结果】分离得到的原始菌株经鉴定为球孢白僵菌(Beauveria bassiana),命名为Beauveria bassiana Bb1003。通过对致死率和正突变率的考察,确定紫外-微波复合诱变的最佳诱变条件为紫外(功率为15 W)照射30 min,微波(功率为800 W、额定微波频率2 450 MHz)辐照60 s。筛选后得到6株复合诱变菌株(UMCM1、UMCM2、UMCM3、UMCM4、UMCM5和UMCM6)。菌株UMCM2对家蚕的僵化率高达...     

北冬虫夏草菌丝体片段诱变育种

目的：通过诱变育种提高北冬虫夏草菌种虫草多糖的含量。方法：对北冬虫夏草子实体进行组织分离,制备菌种,以此作为出发菌株,利用紫外线、微波及其二者结合分别对出发菌株进行诱变,并以其多糖含量为试验指标,探究北冬虫夏草菌丝体片段的诱变育种。结果：通过复合诱变（紫外40s、微波5s,生长正常）的菌株,经测定其多糖含量比对照菌株高17%。结论：通过对北冬虫夏草菌丝体片段进行诱变,找到多糖含量显著提高的菌株是可行的。     

金针菇菌种诱变选育的研究

目的:比较紫外线、微波单因子对金针菇J05Y-C单细胞悬液的诱变效应,并筛选获得优势突变菌株.方法:分别采用不同强度的紫外线、微波单因子对金针菇单细胞悬液进行诱变处理,以出发菌株作为对照,比较诱变致死率、正变率、HC值、菌丝生长量,进行优势菌株筛选、传代稳定性试验.结果:诱变效应的最佳剂量范围在致死率75%～90%的处理组;相同处理时间内,微波诱变的正变率大于紫外诱变的正变率,分别为85.00%和76.67%.采用最大功率700W、脉冲频率2540MHz的微波炉,以中等功率强度处理80s,获得HC值、菌丝生长量比出发菌株提高32.38%、62.16%的优势突变株W8011,连续8次传代遗传性能稳定.结论:在试验的金针菇菌种诱变选育中,微波诱变效应优于紫外线诱变效应.     

枯草芽孢杆菌B-903菌株的诱变选育

枯草芽孢杆菌B-903菌株是由河南省农业科学院植物保护研究所从郑州果园中分离得到,其代谢产生的抗菌物质对多种植物病原真菌具有较强抑制作用。以此菌株为出发菌株,进行亚硝基胍（NTG）和微波诱变处理,确定了,二者诱变处理的最佳处理剂量：NTG最佳处理浓度为200μg／mL,微波诱变为HI微波挡（850W、脉冲频率2450MHz）处理100s,筛选出13个高效突变株。经传代实验,2株高效突变株N1和W2抑菌圈直径分别稳定在26mm和24mm以上,比出发菌株提高21．8％和14．8％．     

ENU诱导点突变——大规模基因突变和功能研究

ENU诱导点突变正在成为一种大规模基因功能研究的有效手段。这里介绍了ENU诱变的机制、影响诱变效率的因素、ENU诱变的策略、表型的筛选、点突变的鉴定以及相关的研究和取得的一些最新进展。     

Mutational approach for N2-fixing and P-solubilizing mutant strains of Klebsiella pneumoniae RSN19 by microwave mutagenesis

Nitrogen (N) fixing Klebsiella pneumoniae RSN19 has high inorganic phosphorus (P) solubilizing capability, but its N₂-fixing capability is limited. In order to acquire a P-solubilizing mutant strain with high efficiency N-fixing capability, different microwave irradiation intensities and durations were tested on RSN19 in an attempt to produce mutants with improved N₂-fixation and P-solubilization capabilities. The effect of microwave irradiation power and time were studied and the microwave mutagenesis parameters were optimized. Nitrogenase activity was tested on the mutant strains by acetylene reduction method; and their P-solubilizing capability and genetic stability were determined. The results indicated that the best conditions for microwave mutagenesis that produced better performed mutant strains were 250W, 36 s. Under these conditions a maximum positive mutation rate of 1.66% was obtained, resulting in five genetically stable strains with promoted nitrogenase activity which was designated as RSM-219, RSM-206, RSM-224, RSM-225 and RSM-275. Subculture tests showed that RSM-219 and RSM-206 were genetically stable mutant strains with higher nitrogenase activity and phosphate solubilizing capabilities than the original strain. Both RSM-219 and RSM-206 performed better than the original strain under N-free conditions when supplied with calcium phosphate only, and produced greater increases in the biomass of alfalfa seedlings.     

T-DNA插入突变在植物功能基因组学中的应用

T-DNA插入突变在植物功能基因组学研究中发挥着重要作用,广泛应用于大规模植物基因功能分析,是分离新基因、研究基因功能的有效工具。我们简单介绍了T-DNA插入突变的原理,详细论述了3种T-DNA插入突变载体的应用,并综述了利用T-DNA插入突变克隆新基因的方法,同时指出了T-DNA插入突变存在的问题及其发展方向。     

微波诱变结合抗性突变筛选放线菌素D高产菌株

采用微波结合链霉素抗性筛选法选育放线菌素D的高产菌株。通过考察链霉素对Streptomyces rubiginosohelvolus FIM-N31菌株孢子生长情况的影响确定链霉素致死浓度,出发菌株FIM-N31的孢子经微波辐照处理后,涂布在含链霉素致死浓度（50 μg/mL）的培养基平板上培养,获得了大量的链霉素抗性基因突变株。摇瓶发酵筛选突变株,结果获得一株遗传性状稳定的放线菌素D高产菌Str186,其产放线菌素D的能力比出发菌株提高了8倍以上。     

常压室温等离子体技术在微生物诱变中的应用进展

微生物是人类赖以生存的重要资源,为提高微生物的生产效率或者赋予其新的生物学功能,需要通过理化方法进行诱变或通过分子生物学技术对其进行定点突变。在目前的理化诱变方法中,常压室温等离子（atmospheric and room temperature plasma,ARTP）诱变技术具有操作简单、条件温和、安全性高、诱变快速等优点,成为倍受青睐的新方法。基于此,综述了ARTP诱变技术的原理及其在微生物诱变育种方面的应用,以期为选育性能优越的微生物菌种的诱变育种相关研究提供借鉴。     

复合诱变选育绿色产色链霉菌抑菌高活性菌株

以一株绿色产色链霉菌xzte06菌株为出发菌株,依次通过软X射线、低能离子束诱变、紫外线照射和微波诱变手段对菌株进行复合诱变,得到一株对产气荚膜梭状芽胞杆菌具有较强抑制活性的菌株W26菌株,且该菌株的遗传特性稳定。对菌株进行抑菌试验发现,该菌株液体深层发酵菌丝体的内酮提取物的抑菌活性与出发菌株相比提高了101.42%。     

插入突变在水稻功能基因组学中的研究进展

构建饱和的基因突变体库是最直接、有效的分析鉴定基因功能的方法.根据插入突变源不同可分为T-DNA插入突变、转座子插入突变等.主要介绍这两种方法的原理及其在水稻功能基因组学研究中的应用和进展,并分析和讨论了插入突变在水稻功能基因组学研究中存在的困难和发展趋势.     

Tests of mutagenesis and reproduction in male rats exposed to 2,450-MHz (CW) microwaves

Tests of mutagenesis and reproduction were conducted in male rats which were irradiated by 2,450-MHz, continuous-wave (CW) microwaves, 4 hr/day from day 6 of gestation to 90 days of age at 5 mW/cm²; or 5 hr/day for five days beginning on the 90th day of age at 10 mW/cm²; or 4 hr/day, 5 days/ wk for four weeks, beginning on the 90th day of age. During selected weekly periods after treatment, the rats were bred to pairs of untreated, normal female rats that were examined in late pregnancy by means of the dominant lethal assay. The reproductive efficiency of these males, as reflected in their breeding, was also examined for changes relating to their microwave experience. No significant evidence of germ-cell mutagenesis was detected when data of microwave-exposed males were compared with those of sham-exposed males, even though there were significant increases in rectal and intra-testicular temperatures at a power density of 28 mW/cm². Temporary sterility, as indexed by fewer pregnancies, was seen at the highest power density.     

Polishing the craft of genetic diversity creation in directed evolution

Genetic diversity creation is a core technology in directed evolution where a high quality mutant library is crucial to its success. Owing to its importance, the technology in genetic diversity creation has seen rapid development over the years and its application has diversified into other fields of scientific research. The advances in molecular cloning and mutagenesis since 2008 were reviewed. Specifically, new cloning techniques were classified based on their principles of complementary overhangs, homologous sequences, overlapping PCR and megaprimers and the advantages, drawbacks and performances of these methods were highlighted. New mutagenesis methods developed for random mutagenesis, focused mutagenesis and DNA recombination were surveyed. The technical requirements of these methods and the mutational spectra were compared and discussed with references to commonly used techniques. The trends of mutant library preparation were summarised. Challenges in genetic diversity creation were discussed with emphases on creating “smart” libraries, controlling the mutagenesis spectrum and specific challenges in each group of mutagenesis methods. An outline of the wider applications of genetic diversity creation includes genome engineering, viral evolution, metagenomics and a study of protein functions. The review ends with an outlook for genetic diversity creation and the prospective developments that can have future impact in this field.