提高普通野生稻花药 培养绿苗诱导率的研究

普通野生稻花培愈伤组织绿苗分化率与材料有很大关系;植株性状较接近栽培稻性状的分化率高;在孕穗前施用复合肥或氮肥的分化率也有明显的提高;接种前稻穗在B—10℃低温预处理8—12天,可提高绿苗分化率1—2倍;愈伤组织诱导以N6培养基为基础,改用MS培养基的有机物,将氨基酸提高到4mg/1,添加6—BA0.5mg/1,丙氨酸2mg/1,诱导率和分化率都高。分化培养基是以MS为基础,把氨基酸和铁盐提高1倍时,绿苗分化率有明显提高。转移到分化培养基的时间应在肉眼见到愈伤组织一周内为好。     

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。     

影响籼粳稻杂交F1花药培养效果的因素分析

以津稻291/IRBB60籼粳杂交F1、反交F1及其亲本的花药为外植体,对诱导和分化培养基、激素配比及有机附加物和高温预培养时间等影响花药培养效果的因素进行研究。结果表明,在SK3基本培养基中添加2 mg/L 2,4-D,1 mg/L NAA和1 mg/L KT比只添加2 mg/L 2,4-D的愈伤诱导率高0.67～2.78个百分点;在诱导培养基中添加水解酪蛋白能明显提高正交F1的愈伤诱导率;MS 1 mg/L NAA 2 mg/L 6-BA 0.5 mg/L KT分化培养基得到了较高的绿苗分化率;正交F1的愈伤诱导率、绿苗分化率和花培力显著高于反交F1;30℃高温预培养36 h,正交F1花培力达到最高值。     

泽林考兰薄层细胞的再生体系

为建立泽林考兰薄层细胞的再生体系,以其试管苗茎尖薄层细胞为外植体,研究了不同激素浓度配比对其类原球茎诱导、增殖、分化及生根的影响。结果表明,茎尖薄层细胞在1/2MS+6-BA 2.0 mg/L+TDZ2.0 mg/L+NAA 0.1mg/L培养基上的类原球茎的诱导率最高,达53.3%;类原球茎在1/2MS+6-BA 2.5 mg/L+NAA 0.1 mg/L培养基上培养40 d后,增殖系数为9.6,类原球茎在1/2MS+6-BA 0.5 mg/L+NAA 0.15 mg/L培养基上培养40 d后分化出小苗,分化率为89.7%;小苗在1/2MS+NAA 0.2 mg/L+AC 0.5 g/L培养基上生根培养40 d后,生根率达94.2%;小苗驯化移栽成活率达90%以上,长势良好。本研究通过以泽林考兰试管苗茎尖薄层细胞为外植体诱导出类原球茎,建立了泽林考兰薄层细胞再生体系,为泽林考兰试管苗产业化生产提供了新的途径。     

S-3307对水稻花药愈伤组织诱导、分化及其壮苗的效应

研究了植物生长延缓剂S-3307对水稻花药培养的效应.在诱导培养基中,低浓度(≤1mg/L)的S-3307可以明显提高愈伤诱导率和绿苗分化率,高浓度(≥10mg/L)的S-3307却致花药发黄,几乎不能生长.在分化培养基中添加低浓度(≤1mg/L)S-3307,有助于绿苗分化率成倍提高,当S-3307的浓度为1mg/L时,绿苗分化率达到最大值,同时白苗分化率显著降     

培养基和培养时间对水稻成熟和胚培养力的影响

本试验研究了10种不同成份的培养基以及延长诱导培养时间对水稻成熟种胚培养力的影响。结果表明,水稻品种对不同培养基的反应是不同的。以∠S—5(∠S基本培养基+1.5mg/L2.4—D+1mg/1 KT+2%蔗糖+0.8%琼脂,pH5.8)培养基的愈伤组织诱导率为最高,诱导的愈伤组织质量较好。诱导的愈伤组织其绿苗分化率也较高,诱导培养60天其愈伤组织仍具有较高的分化能力。     

野生软枣猕猴桃组织培养及褐变处理

以野生软枣猕猴桃嫩芽、幼嫩叶片以及茎段作为外植体,研究野生软枣猕猴桃组织培养整个过程,以建立快速高效的野生软枣猕猴桃再生体系。结果表明：最适宜的诱导叶片、茎段愈伤组织的培养基分别为MS+0.5 mg/L 6-BA+1.0 mg/L NAA和MS+0.5 mg/L 6-BA+0.1 mg/L NAA;诱导出愈伤组织在MS+0.6 mg/L 6-BA+0.05 mg/L NAA培养基上能很好地分化不定芽苗;诱导幼芽产生丛生芽的培养基为MS+1.0 mg/L 6-BA+0.1 mg/L NAA;较适宜的生根培养基为1/2MS。愈伤组织继代中发现细胞生长素NAA可以防止愈伤褐化。在愈伤组织分化中,发现将分化出的芽苗再次愈伤化,可以提高分化率。     

膜荚黄芪愈伤组织诱导及植株再生体系的建立

为了建立膜荚黄芪组培快繁技术体系,本研究以膜荚黄芪无菌试管苗的下胚轴为外植体,探讨不同激素种类及其浓度配比对愈伤组织诱导率、丛生芽诱导率、再生苗生根率的影响,筛选适宜的培养基配方。结果表明:膜荚黄芪愈伤组织最佳诱导培养基是MS+6-BA 0.5 mg/L+2,4-D 2.0 mg/L,愈伤组织诱导率达86.4%,丛生芽分化的最佳培养基是MS+6-BA 1.0 mg/L+NAA 0.3 mg/L,丛生芽分化率最高达91.6%,再生苗最佳生根培养基1/2MS+IBA 0.5 mg/L+IAA 1.0 mg/L,生根率最高达62.6%。本研究建立了膜荚黄芪组培快繁技术体系,为其种质资源保存及开发利用提供了有力的技术支撑。     

藜麦组织培养快速繁殖体系建立研究

为了探讨不同培养基、不同激素组合对藜麦茎段愈伤组织的诱导及分化形成再生植株的影响,以台湾红藜(Chenopodium quinoa Wild.)的茎段为外植体进行诱导、分化增殖、生根、炼苗移栽等研究,建立了藜麦植株再生体系。结果表明:MS+6-BA 1.5mg/L+NAA 0.2mg/L为愈伤组织诱导的最佳培养基,诱导率为84.33%,MS+6-BA 2.0 mg/L+NAA0.6mg/L+2-IP 2mg/L培养基中,愈伤组织分化率最高。     

提高野栽稻杂交后代花药培养率的研究

取普通野生稻与普通栽培稻杂交组合共１０个，研究了不同组合、不同世代、培养基配比及培养时变温处理等与花药培养效率的关系。结果表明：杂交组合间有差异，普通野生稻与粳型稻杂交其后代花培易成绿苗；Ｆ１比Ｆ２、Ｆ３愈伤组织诱导率和分化率均高；培养基以改良Ｎ６培养基最好：变温处理可提高愈伤组织诱导率和绿苗分化率     

白芦笋花药愈伤组织诱导及绿芽分化

为建立芦笋单倍体育种技术体系以解决国内芦笋新品种资源贫乏问题,笔者以白芦笋花药为外植体进行了愈伤组织的诱导及绿芽分化研究。试验结果表明,芦笋花药经过4d的冷藏(4℃)处理后,接种于含有NAA2.0 mg/L、6-BA1.0mg/L及6%蔗糖的1/2MS培养基上,可产生愈伤组织;每年的4月份接种花药的愈伤组织诱导率最高,8月份次之,11月份最低。品种间、同一品种不同单株间花药的愈伤组织诱导率不同,以‘UC155’品种(18.9%)最高,显著高于其他三个品种‘UC142’(11.6%)、‘Gijnlim’(10.3%)、‘Thielim’(5.8%),所有试验株系中以‘UC155’-8的愈伤组织诱导率最高,达83.5%;每日补充光照强度为1000 lx的光照,比全暗培养更有利于芦笋花药愈伤组织的形成;花药接种前进行离心处理(4000 r/min)30 min,不能提高愈伤组织诱导率;愈伤组织转瓶培养于NAA 0.5 mg/L、6-BA 1.0 mg/L及3%蔗糖的MS培养基上,能分化出无根绿芽;不同品种间绿芽分化率有差别,以‘Gijnlim’品种最高(67.2%),‘UC142’品种最低(14.1%);未分化绿芽的愈伤组织再转入于不含植物生长调节剂的MS培养基上培养,部分愈伤组织能分化出绿芽     

水稻高效花药培养技术体系的构建

为提高水稻花药培养效果,产生大量的花粉植株,以多年的试验并结合多方面的研究结果,论述了水稻花药培养各个环节中的关键技术,创建了通常情况下的一套水稻高效花药培养技术体系。本系统着重于在利用基因型的选择和基本配养基的交叉使用以提高花药愈伤组织诱导率的的前提下,在其它的每个环节,尤其是壮苗和移植管理技术方面进行了优化改进,以提高绿苗分化率和组培苗移植后的成活率。     

天蓝苜蓿花药愈伤组织诱导及再生体系的建立

为获得天蓝苜蓿单倍体再生植株,本研究以野生天蓝苜蓿花药为外植体,采用正交设计L₁₆(4⁴)筛选适宜天蓝苜蓿愈伤组织诱导培养基,比较不同基本培养基及生长调节剂组合筛选适宜的分化培养基,并用1/3MS、1/2MS和MS添加不同浓度的NAA研究不定生根。结果表明:天蓝苜蓿现蕾15~25 d的花药其愈伤组织诱导效果最好,高达60.5%。4℃低温预处理2~4 d有利于愈伤组织诱导,诱导率达77.2%。适宜花药愈伤组织诱导的培养基为NB+2,4-D 1.0 mg/L+NAA 0.5 mg/L+6-BA 0.5 mg/L+TDZ 1.0 mg/L,诱导率达78.5%。比较不同基本培养及生长调节剂的不同组合发现,NB培养基愈伤组织分化效果优于B₅和MS,NB+NAA0.5 mg/L+6-BA 2.0 mg/L适宜愈伤组织的分化,分化率为66.3%。不定芽在1/3MS+NAA 0.5 mg/L中培养,生根率最高,为86.55%。本研究建立了天蓝苜蓿花药培养再生体系,获得了单倍体植株,为天蓝苜蓿的育种实践及基因组学研究提供基础材料。     

Enhanced anther culture efficiency of indica rice (Oryza sativa L.) through modification of the culture media

The production of microspore-derived green plants from anther culture of indica rice is generally very low compared with japonica cultivars. A modified anther culture medium, consisting of a higher KNO₃ content (31 mm ) and casein hydrolysate (CH, 500 mg/1) but without ammonium salts, was tested in comparison with a medium consisting of the widely-used N6 medium nitrogen background, using four indica × indica F₁ hybrids as test materials. Green plant regeneration frequency was at least three-fold higher in the microspore-calli derived from the former medium than in those derived from the modified N6 medium. More than 700 microspore-derived plants were raised in the field. Another study was carried out using indica × japonica and indica × javanica F₁ hybrids. The results indicated that a medium with higher (3.5 mm ) ammonium sulphate may induce a higher frequency of anthers with microspore-calli but not necessarily lead to a larger number of green-plant regenerating calli. Subsequently, using the indica cv. ‘IR-43’ as the test material, use of a lower level (1.75 mm ) of (NH₄)₂SO₄, in addition to KNO₃ (31 mm ), was found to be better than CH (500 mg/l) for anther-response as well as green plant regenerability of the derived microspore-calli. Nitrate-nitrogen or ammonium-nitrogen alone elicited poor response. Twenty-five media involving combinations of KNO₃ (20–34 mm ) and (NH₄)₂SO₄ (1–3 mm ) were tested for their effects on anther response. Combinations involving KNO₃ (31–34 mM) and (NH₄)₂SO₄ (2.0–2.5 mm ) were found superior not only for achieving greater anther response but also, for subsequent green-plant regeneration. This contrasts with the 28 mm of KNO₃ and 3.5 mm of (NH₄)₂SO₄ in the widely-used N6 medium developed for japonica rice. Other than potassium nitrate and ammonium sulphate, and potassium phosphate to some extent, the levels of other inorganic salts tested did not make any significant difference to the process of anther response. Based on these results, modified media with three levels of ammonium sulphate were tested for anther culture efficiency of indica × japonica and indica × javanica derivatives (F₃s). Microspore-calli derived from a medium of a lower (1.75 mm ) level of (NH₄)₂SO₄ showed a higher regeneration potential overall than those derived from a higher (3.5 mm ) level. A revised medium has been suggested, on the basis of these results, for the realization of improved anther culture efficiency and, consequently, improved feasibility of using doubled haploids in genetic and breeding research with indica rice.     

水稻籼粳交偏籼后代材料花药培养研究

试验利用水稻籼粳交偏籼后代的10份材料进行了花药培养效果研究,结果如下:不同株系间培养效果存在较大差异,出愈率和绿苗分化率之间没有必然联系;N6、SK3培养基对籼粳交偏籼后代材料的培养效果明显优于通用培养基,N6又稍优于SK3培养基;花培材料在预冷处理之前不宜剪叶;从试验中筛选到了2份具有较高培养力的株系,为今后遗传育种工作和分子遗传研究提供了合适的遗传工程材料。     

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。     

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。     

提高水稻同源四倍体花药培养愈伤诱导率的研究

以同源四倍体水稻原种零轮、02428、培矮64、轮回422及其成对二倍体为实验材料,分别以琼脂浓度（0.5%、0.75%和1%）、碳源种类（蔗糖和麦芽糖）与用量（60 g L^-1和80 g L^-1）、不同激动素（KT和Zip）以及不同基本诱导培养基（MS、N6、M8、NB）等因素为变量,共设计26种诱导培养基（2种对照）,进行了花药愈伤诱导实验,旨在筛选出愈伤诱导率的培养基优化组合。结果表明,琼脂浓度0.75%和60 g L^-1蔗糖组合对提高水稻花药诱导率作用较好, N6培养基及其衍生NB培养基对提高四倍体水稻特别是粳稻花药诱导率有着较好作用。激动素的加入,能显著提高愈伤诱导率,最高可达到84%。在供试的26种培养基中,有6种诱导培养基得到较高的四倍体粳稻愈伤诱导率,有2种诱导培养基得到较高的四倍体籼稻愈伤诱导率,在8种较好的诱导培养基中有3种不但有较高的愈伤诱导率而且具有较高的绿苗率。     

Anther culture of recalcitrant indica × Basmati rice hybrids

Fertile, green, di-haploid plants were obtained at high frequencies from several indica × Basmati rice F₁ hybrids and/or F₂ plant populations using an improved anther culture procedure. Anthers from cold-pretreated (10 ^°C for 10 d) panicles of six indica (HKR120, HKR86-3, HKR86-217, PR106, Gobind andCH2 double dwarf) and two Basmati rice (Basmati 370,Taraori Basmati) varieties and 14 heterotic indica ×Basmati F₁/F₂ hybrids were cultured in modified agarose-solidified N6M, Heh5M and RZM media. Best callus induction frequencies (2.6–78%) were obtained in RZM medium containing 4% (w/v) maltose,2,4-D, NAA and kinetin. F₂ plants compared to F₁ hybrids and parental rice varieties, were more responsive to anther culture. Androgenesis frequencies of 31–78% were obtained for indica × Basmati F₂ plants in RZM medium in just 30 d which are comparable to or higher than that reported for japonica rice varieties and hybrids involving japonica rice parent(s). Agarose (1.0% w/v)-solidified MS medium containing 3.0% maltose, kinetin, BAP, and NAA, induced green shoot regeneration in 0–51% of the anther-derived callide pending upon the genotype. High plant regeneration frequencies (67–337 green plants per 1000 anthers)were obtained from anther calli of several F₁hybrids (Gobind × Basmati 370 and HKR120 ×Taraori Basmati) and F₂ plants (Gobind × Basmati370, Gobind × Taraori Basmati, HKR86-3 × TaraoriBasmati). A sample of 498 plants obtained from the above hybrids, were transferred to pots with>90% survival; 8–78% of these plants had >5%spikelet fertility and were diploid. In addition,18% of the haploid plants could be diploidized by submerging in 0.1% colchicine solution for 16–18 h. The improved anther culture procedure reported here, resulted in several fold increase in the recovery of green plants from recalcitrant indica × Basmati rice hybrids compared to previous published procedures. The study may accelerate the introgression of desirable genes from indica into Basmati rice using anther culture as a breeding tool. This revised version was published online in July 2006 with corrections to the Cover Date.