双亲灭活原生质体融合技术在草菇耐低温菌株选育上的应用

为了选育耐低温草菇菌株,以草菇V23、V3552为亲本,采用酶解法制备原生质体,用紫外诱变、化学诱变两种方法对草菇菌株进行诱变,筛选到耐低温的突变株;然后利用紫外灭活（20 W,30 cm,110 s）和热灭活（50℃ 3 min）的双亲灭活标记法对突变株进行化学融合,结果表明在400 g/L的PEG6000、pH 8.0、融合时间30 min和融合温度32℃的条件下融合率最高,达到0.517%, 共获得200个融合子。经过0℃低温筛选,最终获得15株草菇耐低温菌株,菌丝在0℃的耐冻能力提高了4.5倍。经出菇实验证明其子实体与出发菌株相比具有明显的耐低温性,液化现象明显推迟,说明该方法筛选出的菌株具有进一步应用开发的价值。     

大白菜突变体库的构建及M2叶片表型变异的研究

 利用不同浓度甲基磺酸乙酯（EMS）诱变大白菜自交系‘A03’种子,根据M₁代植株发芽率、成活率、育性及M₂代成活率,筛选出适宜的EMS诱变浓度为0.4%。采取EMS种子诱变1次、EMS种子诱变2次及EMS花蕾诱变结合小孢子培养的方法构建了含有4 253个家系及相应自交M₂代种子的大白菜突变体库。在该突变体库M₂群体中分别对苗期6 404株幼苗及莲座期7 756个单株的叶片性状进行了形态学调查,总的表型变异频率高达37.62%和26.33%。     

拮抗细菌Enterobacter cloaeae B8的一株抗菌素抗性突变体(英文)

为了研究拮抗细菌Enterobacter cloacae B8在野外水稻叶面上的定殖情况及拮抗作用,由自然选择和Tn5诱变从B8产生了一株抗菌素抗性突变体BX8.BX8能在含利福平达400μg/ml或卡那霉素达300μg/ml的LB培养基中生长.实验表明该抗菌素抗性的产生没有降低BX8的拮抗活性.该抗性比较稳定,在无抗菌素培养基中培养.BX8至少分裂生长60代内抗性不变.     

植物离体诱变及在百合育种中的应用

植物离体诱变技术目前已广泛应用于作物优质、高产、抗病、抗逆等新种质的筛选中。综述了体细胞无性系变异简况、离体诱变育种采用的技术、抗性突变体的筛选技术以及检测方法、应用现状及在百合育种中的应用情况。分析了离体诱变育种的优势及在百合抗病育种中的应用前景。     

产甲壳素酶菌株的筛选与诱变

以胶体甲壳素作为惟一碳源,从自然界微生物中筛选并诱导到一株产甲壳素酶活性较高的菌株,经16S r DNA鉴定为Aeromonas sp.。并以该菌株作为出发菌株,经过一次紫外诱变,酶活是初筛菌株的1.48倍。再经过二次紫外诱变或者紫外-Li Cl复合诱变,酶活分别是初筛菌株的3.16和3.87倍,最高酶活达到3.48 U/m L。最后通过连续传代培养,确定了菌株HMX-16经过紫外和复合诱变后突变性状稳定性良好。     

紫外诱变吸水链霉菌选育高产农用抗生素菌株

[目的]为抗生素产生菌在农业生产中的应用奠定基础。[方法]以小麦赤霉菌为指示菌,以从吸水链霉菌海南变种S101菌株分离到的S510菌株为出发菌株,经紫外诱变后分别采用琼脂块法和二级摇瓶复筛法对其进行初筛和复筛,以选育出高产菌株,分析发酵培养基pH值和培养时间对发酵液效价的影响。[结果]当发酵培养基的pH值为5.5~6.5时,发酵液效价较高,最适pH值为6.0。发酵液的效价在72 h后可维持在较稳定的范围内,最佳发酵时间为96 h。通过连续2次紫外诱变,菌株的产抗生素能力比初始菌株提高了30.06%。将获得菌株在斜面培养基上连续传代5次后,仍有稳定的遗传性状。[结论]采用紫外诱变法选育该链霉菌是有效的。     

一株好氧反硝化细菌的筛选、鉴定及紫外诱变

从牛蒡(Arctium lappa L.)根际土壤中分离到1株具有较高反硝化能力的好氧细菌YB000,对该菌株采用生理生化及分子生物学方法进行了鉴定,并且对该菌株进行了以提高反硝化性能为目的的紫外诱变。结果表明,分离自牛蒡根际的反硝化细菌经鉴定为铜绿假单胞菌(Pseudomonas aeruginosa),该菌株于距离30 W紫外灯30 cm处照射240 s可获得具有较强脱氮能力且遗传性状稳定的诱变菌株YB004和YB005,其脱氮能力分别达到93.43%、92.03%。     

Inheritance and potentials of a mutated dwarfing gene ndf1 in Brassica napus

A dwarf mutant ‘NDF‐1′, approximately 70 cm high, was derived from a 200‐cm high doubled haploid (DH) line ‘3529’ (Brassica napus), seeds of which were jointly treated with chemical inducers and bombardment of fast neutron. The leaves of the ‘NDF‐1’ mutant were wrinkled and thicker compared with the wild‐type control. The mutant had much lower values than its original parents for all agronomic traits, except for its seed weight. A genetic analysis revealed that dwarfism is under the control of a major gene (designated as ndf1) with a mainly additive effect and non‐significant dominance effect. Because of the high level of resistance to lodging, breeding programmes for double low dwarf oilseed rape and heterosis utilization were initiated. Some new dwarf strains with improved agronomic performance were developed. The hybrid of the cross between the tall parent and the dwarf line showed increased harvest index and significantly higher seed yield than the tall parent or the control variety ‘Zhongyou 821’ and presented an estimated heterosis vigour rate as high as 12.5–25.8%. The dwarf trait will be a promising marker for a simple, economic and efficient way to control the purity of F₁ hybrid varieties in hybrid production of B. napus.     

Inheritance of fiber quality and lint yield in a chemically mutated population of cotton

The narrow germplasm base of the upland cotton (Gossypium hirsutum L.), grown on the Texas high plains historically, has limited improvement of fiber quality. Chemical mutagenesis and subsequent selection have helped the development of lines with improved fiber quality in cultivars adapted to this region. This study was conducted to determine the inheritance of improvements in fiber quality. M₃ lines with divergent fiber properties of micronaire, length, and strength were selected from a population of Paymaster HS 200 treated with 3% v/v ethyl methanesulfonate (EMS) for two hours. The 115 selected lines of M₄ and M₅ generation were evaluated for fiber quality and lint yield. Regression of the M₄ and M₅ on the M₃ generation, as well as the M₅ on the M₄ was used to generate narrow sense heritability coefficients. Significant variations were observed between the mutant lines in all generations except for lint yield in the M₅ (1997). The highest heritability estimates were found in fiber length (h ²= 0.29** to 0.46**). Micronaire and strength showed intermediate heritability estimates of h ²= 0.14 to 0.19, while lint yield had a very low heritability estimate of h ²= 0.03. Fiber length and strength were correlated (r= 0.58** to 0.46**) in all the three generations. The mutants identified in these studies have the potential to improve fiber quality of upland cotton without introducing alien genes that may reduce adaptation to short growing season production regions.     

TILLING and Ecotilling the Plant Kingdom

The modem crop scientist has a large amount of available nucleotide sequence information to identify genes of potential agronomic importance. Using reverse genetic approaches, specific genes can be disrupted, and hypotheses regarding gene function directly tested in vivo. Although a number of reverse genetic methods have been introduced, many are limited in application because they are organism-specific, expensive, transgenic, or only transiently disrupt gene function. However, traditional mutagenesis using chemical mutagens has been widely used as a forward genetics strategy to create many new crop plant varieties at relatively low cost. Mutagens such as ethyl methanesulphonate （EMS） cause stable point mutations and thus produce an allelic series of truncation and missense changes that can provide a range of phenotypes （Greene et al., 2003）. TILLING （targeting induced local lesions IN genomes） is a high-throughput reverse genetic strategy that combines traditional mutagenesis and SNP discovery methods （Colbert et al., 2001; McCallum et al., 2000）. To identify mutations, target regions of-l.5 kb are amplified with fluorescently labeled gene specific primers. Heteroduplexes are then formed between wild-type and mutant strands, mismatches are cleaved by incubation with a single-strand specific nuclease,