水稻花药培养技术的研究进展

花药培养技术的出现,大大地缩短了水稻育种的年限,也加快了各种基因型材料的获取。本研究综述了花药培养技术概况、影响花药培养力的6个关键因素、水稻花药培养技术的进展、分析水稻花药培养技术现阶段存在的问题,并对未来发展进行了展望。     

水稻花药培养及其在遗传育种上的应用

花药培养可以获得单倍体植株,单倍体在植物遗传育种中具有重要意义.水稻花药培养可以快速纯合育种材料、提高选择效率、有效缩短育种周期,扩大变异范围、加速有效性状转移.本文综述了水稻花药培养在基因型选择、取样的低温预处理、培养基配制、接种和培养、分苗和炼苗等技术要点,在常规稻育种、杂交稻育种、水稻基因工程育种等方面应用的主要成果,并对建立高效花药培养技术体系和在水稻遗传育种应用上向宽范围和深层次发展进行展望.     

不同基因型水稻花药培养效果的比较研究

为了筛选到花药培养效果好的水稻材料,选取常规籼稻、籼粳杂交稻、籼型杂交稻和粳型杂交稻 4 种不同类型的水稻材料进行花药培养,比较它们在花药培养过程中的愈伤组织诱导率和花药培养效果,建立高效花药培养再生体系。结果表明,禾香 1 号和福稻 99 愈伤组织诱导率较高,分别为 3.01% 和 1.55%,可作为水稻花培育种的优良亲本。以玉晶 91 为亲本配制杂交组合并进行花药培养,结果表明玉晶 91 与某些亲本间具有较好的花药培养配合力和杂种优势,在水稻花培育种中具有一定的潜在利用价值。粳型材料中,甬优 1526、组合圣稻 18-15// 春江 041/ 武运粳 24、中浙粳 18/ 鄂香 2 号和香 1 号 /17J42 均不适合作为水稻花药培养育种的材料。甬优 2640 的花药培养愈伤组织诱导率和绿苗分化率均较高,可以在水稻花培育种和DH 群体创建中加以利用。这对创制新的种质资源、构建 DH 群体和培育新品种具有十分重要的意义。     

提高水稻花药培养效率分析

提高水稻花药培养效率,一直是科技工作者进行研究的主题。采用低温处理、2,4-D浸茎节（或热处理）、人工加倍、培育壮苗的操作方法,使绿苗生产率由3%～5%提高到8%～14%。提高绿苗生产率1～2倍。以结实苗数统计花药培养效率由0.5%提高到4%～6%,是未采用此技术的8～10倍。     

水稻花药培养研究与应用进展

该文对水稻花药培养研究与应用的历史进行回顾；介绍水稻花药培养技术在粳常规育种、灿稻常规育种、杂交稻中及水稻基因工程育种等方面应用的最新进展；探讨水稻花药培养研究与应用的发展趋势。     

适宜寒地稻区种植的3个水稻花培新品种

自日本人新关和大野通过花药培养获得水稻单倍体植株后,花药培养作为一项新技术受到世界各国科研工作者的重视并在育种上得到有效利用。我国于1970年开展水稻花药培养技术研究,在新材料创造、新品种选育、基因定位和加速转基因水稻外源基因纯合等方面都起到重要的促进作用,并取得了良好效果．使我国粳稻花培育种应用技术研究处于国际领先地位。     

水稻花药培养技术及其研究进展

水稻花药培养是水稻育种中重要的技术手段。本文从材料基因型、取材时期与低温预处理、培养基种类、激素配比、碳源、其他添加剂、培养条件等方面总结了水稻花药培养技术的研究进展,分析了现阶段花药培养存在的问题,并对未来的研究方向做出了展望。     

化学杀雄对籼稻花药培养力的影响

自1968年新关首先获得粳稻花粉植株以来,由于培养技术的不断改进,高效培养基的研制,使粳稻花药培养力大幅度提高,世界各国和一些国际研究组织相继采用花药培养技术应用于水稻育种实践。但迄今籼稻花药培养力仍极低(0.5%左右),致使该技术难以在籼稻育种上应用,而籼稻又在水稻生产中占主导地位,因此籼稻花药培养力的研究仍被列     

提高寒地水稻花药培养效率的几个关键技术

论述了寒地水稻花药培养操作规程中应注意的几个关键环节,以便使该项技术更好地应用于水稻育种,提高工作效率。①根据器官形态指标外推法选择花粉发育处于单核靠边期的幼穗,进行6~8℃低温预处理12d左右,可提高愈伤组织诱导率;②愈伤组织转移大小以直径1~2mm为宜,当绿苗长至4cm左右时及时进行壮苗培养;③做好接种材料、室内、人员的消毒和卫生工作,降低污染率,加强温室等的管理。     

液体培养基在水稻花药培养中的应用研究

采用两种培养方式（液体培养基诱导法、固体培养基诱导法）对水稻花药进行培养,结果表明,液体培养基诱导法与固体培养基诱导法培养时间相当,但液体培养基诱导法的出愈率及绿苗分化率均提高10%左右,由此认为,液体培养基诱导法是提高水稻花药培养效率的可行方法。     

小麦花药培养的基因型差异与亲本选配分析

对104份不同基因型材料进行花药培养,结果表明：F1代愈伤组织诱导率为13.28%,高于F3代的6.02%; 绿苗产率F1代为2.88%,F3代为1.10%,F1相当于F3的2.6倍。同样的培养条件下,不同基因型材料间花药培养力差异很大,愈伤诱导率在 0～111.43% 之间、绿苗产率在 0～49.29% 之间;愈伤诱导率、绿苗分化率与绿苗产率三者之间成正相关关系。同时筛选出了一批如宁春4号等具有高培养力、高产和优质基因的花培桥梁亲本,为有目的配制杂交组合提供依据。     

Anther culture response of wild Oryza species

Thirteen different wild species of the genus Oryza were investigated for their response to anther culture and plant regeneration. Callus induction from microspores of anthers was found to be strongly dependent on the species. Although large numbers of anthers from wild Oryza species, including O. barthii, O. latifolia, O. australiensis, O. minuta, O. nivara, O. paraguagensis and O. eichingeri, were cultured, no calli could be obtained. However, calli were produced from anthers of O. punctata, O. perennis, O. alta, O. ridleyi and O. rufipogon, although the frequency of callus induction was different. Similar species-dependence was observed in plant regeneration from microspore-derived calli. In total, 62 plants were derived from anther culture, including 47 albino and 15 green plants (of which 26.7% were haploids) from O. perennis; for the first time, six albino plants were obtained from O. ridleyi. Phytohormone combinations in the callus induction medium were found to influence callus induction and different wild Oryza species exhibited their own preferred phytohormone combinations for anther culture. In general, NK medium containing suitable concentrations of auxin and cytokinin may be successfully applied for anther culture of selected wild Oryza species.     

Anther culture in connection with induced mutations for rice improvement

Doubled haploids have long been recognized as a valuable tool in plant breeding since it not only offers the quickest method of advancing heterozygous breeding lines to homozygosity, but also increases the selection efficiency over conventional procedures due to better discrimination between genotypes within any one generation. Ten cultivars of japonica rice and nine cultivars of indica rice were evaluated for androgenic response. Various doses (10–50 Gy) of gamma rays were applied to investigate the effect of radiation on callus formation, green plant regeneration and the frequency of selected doubled haploid mutants. Similarly, the effects of colchicine concentration (10–200 mg/l) on callus induction, regeneration and fertility of green plants were observed. It was demonstrated that the dose of 20 Gy gamma rays and 30 mg/l concentration of colchicine have significant stimulation effect on regeneration of green plants from rice anther culture. The high frequency of observed doubled haploid mutants indicates that anther culture applied in connection with gamma rays is an effective way to improve rice cultivars. This revised version was published online in August 2006 with corrections to the Cover Date.     

Somatic in vitro culture response of Lolium perenne L.: genetic effects and correlations with anther culture

Summary Genotypic effects on callus induction and plant regeneration in callus, suspension and protoplast culture, and their correlations with both phenotypic and GCA-values for anther culture response, were studied using 21 genotypes of perennial ryegrass. Differences between genotypes accounted for approximately 40% of the total variation for callus induction and initial callus growth, and 59 and 83% of the variation in callus culture for regeneration percentage and percentage of green plants. Effects of genotypes were less pronounced in suspension culture, where suspensions from the same genotype often behaved differently. Some suspension cultures retained their capacity for green plant regeneration for almost two years, repeatedly producing 80–100% green regenerants during this period. Genotypes with high regeneration percentage and a large proportion of green plants from callus culture were also superior in suspension culture for both regeneration performance and longevity. Regeneration percentage and percentage of green plants were uncorrelated, and probably under different genetic control. While capacity for green plant formation from the different genotypes showed no correlation between anther culture and somatic in vitro culture, a positive correlation was observed between the regeneration percentages in somatic in vitro culture and anther culture (r=0.44^*–0.85^***), suggesting some common genetic control of the two systems.     

光身稻的培养特性研究

本文对光身稻培养特性进行了系统研究。结果指出:(1)光身稻的成熟胚培养力极低，其绿苗分化率几乎等于零，为0.7%，但其幼穗培养力则较高，绿苗分化率为44.8%; (2)光身稻的花药愈伤组织出现时间较早，为25天左右，而且愈伤组织诱导率也较高，为7%，但其绿苗分化率很低，仅为0.35%; (3)光身稻的适宜诱导培养基为M8; (4)花药愈     

基因型对陆地棉花药离体培养反应的影响

对２７种棉花基因型的花药在离体培养中的反应进行了比较,结果表明：所有的基因型在合适的培养条件下均能高频率地形成愈伤组织,但不同基因型间愈伤组织诱导率和诱导量不同;再生植株的花药比原始亲本的花药易诱导出愈伤组织;品种间杂种表现出部分优势,但不明显。本试验中,仅鲁棉６号和Ｓｉｏｋｒａ１－３两个品种诱导获得了胚性愈伤组织、胚状体和再生植株,且二者之间也存在着差别。从基因型在棉花花药培养中的效应、外界调控对棉花不同基因型花药培养效果的影响、棉花花药培养与体细胞培养基因型差异的比较等三个方面进行了讨论,提出了棉花花药培养愈伤组织诱导和植株再生是受两套遗传基因控制的遗传性状,并提出了棉花组织培养植株再生的阈值问题     

水稻愈伤组织生长速率研究

通过对水稻愈伤组织生长速率的研究,发现水稻愈伤组织生长速率因培养条件的不同和外质体的不同有很大差异,培养温度对愈伤组织生长速率影响较大,较小的温度降低可以使水稻愈伤组织生长速率降低很多,水稻愈伤组织生长速率与连续继代培养时是否改变2,4-D浓度有一定关系,一般表现为在较低温度下改变2,4-D浓度进行继代,可以促进水稻愈伤组织生长;在较高温度下改变2,4-D浓度进行继代,对水稻愈伤组织生长速率影响不大。在23~25℃培养条件下,“中花11”水稻花药培养的单倍体愈伤组织,比其成熟胚诱导的二倍体愈伤组织生长速率快一倍左右。继代时的初始接种量的多少对水稻愈伤组织生长速率也有关系。     

不同世代太谷核不育小麦对花药成株的影响

通过对太谷核不育小麦杂交一代可育株和不育株,及二、三、四代可育株花药的离体培养,结果表明:1.不育株花药培养能够获得植株再生;2.杂交一代的可育株花药出愈率最高,绿苗分化率亦最高;3.基因型对花药出愈率和绿苗分化率有明显的影响.     

5个酿酒葡萄品种组织培养及再生体系的建立

为了建立酿酒葡萄离体培养及植株高效再生体系,以5个酿酒葡萄品种花药和茎尖为外植体,利用组织培养法,研究了外源激素、基因型对外植体培养及再生的影响。结果表明,茎尖愈伤组织诱导及分化均优于花药。茎尖愈伤组织诱导培养基为：B5+NAA 0.5 mg/L+6-BA 1.0~2.0 mg/L;在此培养基上添加AgNO3 10 mg/L或PVP 1000 mg/L对花药愈伤组织诱导较好。茎尖愈伤组织分化培养基为：B5+NAA 0.01 mg/L+6-BA 0.5 mg/L+GA3 0.2 mg/L,最高分化率达100%。本试验成功地建立了酿酒葡萄茎尖愈伤组织诱导、再分化芽苗再生体系,该结果可为酿酒葡萄良种快繁以及遗传转化体系建立奠定良好的基础。     

宁夏优质水稻品种D10高效再生体系的建立

为了利用基因沉默技术定向改良宁夏香稻品种,短期内建立可获得成苗的高效成熟胚再生体系,以宁夏优质水稻品种D10成熟胚作为外植体,通过植物组织培养的方法,研究了培养基、外源激素、光照和温度对外植体培养及再生的影响。结果表明：不同培养基对愈伤组织诱导率有着不同的影响,MS培养基最低,N6D培养基最高;32℃持续光照培养的愈伤组织明显优于30℃ 12 h/d光照培养;在分化培养基中添加TDZ,发现浓度为1 mg/L的TDZ与低浓度的IAA结合使绿苗分化率提高到100%;另外添加2 mg/L ABA的N6D培养基中愈伤组织诱导率为100%。最终该品种的最优再生体系为：32℃持续光照培养并添加ABA 2 mg/L的N6D培养基;分化培养基为B5+TDZ 1mg/L+IAA 0.2mg/L;生根培养基为1/2MS+KT 2 mg/L+NAA 0.2 mg/L+CH 2 g/L+Sorb 30 g/L。本研究建立了宁夏优质水稻品种D10成熟胚再生体系。