Effects of Physiological Status of Parent Plants and Culture Medium Composition on the Anther Culture of Sorghum

Abstract

Anthers at various maturation stages were collected from seven genotypes of Sorghum bicolor, and cultured on MS medium supplemented with 3 mg L^?1 2,4-D and 2.0 mg L^?1 kinetin. The rate of callus induction was high in young anthers, and decreased with maturation of the anther. Young anthers were collected from the plants grown in the greenhouse (greenhouse anthers), in the field (field anthers), and in the chambers kept at 20, 25 or 30 °C. Then they were cultured on MS medium containing 2.0 mg L^?1 kinetin, 1.0 mg L^?1 IAA and 2.5 mg L^?1 2,4-D, on which the rate of callus induction was the highest. The average callus-induction percentage was 6.4% in the field anthers, and 3.7% in the greenhouse anthers. Callus induction was low in the anthers collected from the plants grown at 30° C. Among the genotypes examined, TX403 produced calli at the highest frequency. The rate of plant regeneration from anther-calli was the highest on the medium containing 2.5 mg L^?1 kinetin and 3.0 mg L^?1 IAA. The regeneration rate of green plantlets on this medium was 27% on the average, and was 46.2 and 28.6% in PP290 and TX403, respectively. The chromosome number in root-tip cells was unstable, but was 20 in most of the regenerated plantlets.     

低浓度2,4-D提高水稻体细胞成苗研究

 以较低浓度(0.2、0.4、0.8 mg/L)的2,4-D作为水稻(Oryza sativa L.)的外植体(种子)诱导愈伤组织培养基中的生长素类激素，发现0.8 mg/L的2,4-D能较早地使胚的盾片产生大量的愈伤组织，但这些愈伤组织在原培养基上不能分化出绿芽。含0.2 mg/L的2,4-D能在胚芽鞘节等部位产生一些较光滑、致密、白色颗粒状的结构或愈伤组织，虽然它们出现较迟，但这些结构物能在原诱导培养基上逐渐转化成芽簇，并产生一些纤细的根。将芽簇分割成小块，转移到加有KT等的成苗培养基上，能形成大量的幼苗，绿苗频率达80%以上，明显高于一般报道的水平。试管苗经大田种植后生长整齐一致，能产生出正常的种子。     

云蔗03-194甘蔗组培快繁技术

以甘蔗新品种云蔗03-194的幼嫩叶片作为外植体,采用不同2,4-D浓度对幼嫩叶片的切片进行愈伤诱导,以不同6-BA和KT激素配比对甘蔗愈伤进行分化诱导和增殖培养,以不同NAA浓度及香蕉汁添加量诱导甘蔗组培苗生根。结果表明:适宜甘蔗幼嫩叶片愈伤诱导的培养基为MS+2,4-D 1.5mg/L,诱导分化培养以MS+6-BA1mg/L+KT 0.5 mg/L为宜,增殖培养以6-BA 2.0 mg/L+KT 1.0 mg/L为宜,适宜的生根培养基为MS+NAA 1.5mg/L+30mL香蕉汁。以河沙:红壤土(2∶1)为假植基质,假植成活率达92%～96%,植株生长健壮,长势良好。     

低浓度2，4－D提高水稻体细胞成苗研究

以较低浓度（０．２、０．４、０．８ｍｇ／Ｌ）的２，４－Ｄ作为水稻（ＯｒｙｚａｓａｔｉｖａＬ．）的外植体（种子）诱导愈伤组织培养基中的生长素类激素，发现０．８ｍｇ／Ｌ的２，４－Ｄ能较早地使胚的盾片产生大量的愈伤组织，但这些愈伤组织在原培养基上不能分化出绿芽。含０．２ｍｇ／Ｌ的２，４－Ｄ能在胚芽鞘节等部位产生一些较光滑、致密、白色颗粒状的结构或愈伤组织，虽然它们出现较迟，但这些结构物能在原诱导培养基上逐渐转化成芽簇，并产生一些纤细的根。将芽簇分割成小块，转移到加有ＫＴ等的成苗培养基上，能形成大量的幼苗，绿苗频率达８０％以上，明显高于一般报道的水平。试管苗经大田种植后生长整齐一致，能产生出正常的种子。     

苯乙酸促进水稻花药愈伤组织的再分化和直接成苗

 研究了苯乙酸（phenylacetic acid，PAA）应用于水稻花药培养的效果。以 2,4-D 为对照，发现PAA不影响愈伤组织诱导率，但显著提高了愈伤组织分化率（p<0. 05），从而显著提高了植株再生率（p<0. 05）。其促进效果与品种、培养基无机盐成分有关，对籼稻品种效果较好。PAA还能有效地促进花药愈伤组织不经转移到分化培养基上而直接在诱导培养基上分化成苗。直接再生的绿苗大多具有旺盛的不定根，可直接移栽到土壤中去。当PAA浓度为10 mg/L 时，特青、02428和亚优2号的直接植株再生率分别达0. 56%, 1. 64%和2. 69%。提高PAA浓度可进一步提高直接植株再生率。利用基于PAA的一步成苗培养法已构建成了3个水稻杂种花培后代加倍单倍体（DH）群体。     

Efficient Plant Regeneration in Garlic through Somatic Embryogenesis from Root Tip Expiants

Abstract

The present study was conducted to establish an efficient protocol of plantlet regeneration through somatic embryogenesis in garlic (Allium sativum L.). Root tips measuring 2 to 3 mm were excised and cultured on agar-solidified MS medium containing various concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) for callus and embryo formation. The optimum concentration of 2,4-D was 0.5 μM. At a concentration higher than 1.0 μM, 2,4-D had an inhibitory effect on callus and embryo formation. Embryos germinated and formed rooted plantlets on MS solid medium containing 5.0 μM kinetin. The number of plantlets regenerated per root tip expiant depended on the concentration of 2,4-D in the callus initiation medium. The plantlets were established in the soil after acclimatization in a growth cabinet. Somatic embryos were morphologically characterized by scanning electron microscopy (SEM).     

荔枝"妃子笑"品种花药培养及其体胚发生

以荔枝(LitchichinesisSonn.)品种“妃子笑”为试材,研究其花药离体培养及植株再生的影响因素。结果表明,荔枝花药在MS 2,4-D2mg/L NAA0.2mg/L以及含50g/L蔗糖的培养基上诱导胚性愈伤组织效果较好,把胚性愈伤组织转移到MS BA1mg/L NAA0.5mg/L,谷氨酰胺500mg/L,蔗糖50g/L分化培养基上培养1个月后,体胚大量萌发,再将成熟体胚转移到附加500mg/L谷氨酰胺的MS无激素培养基上,培养1￣2个月后,能再生成完整植株。     

冬小麦高花药培养力基因型的筛选

为了筛选小麦花培育种骨干亲本,减轻花培育种的基因型依赖性问题,对74个冬小麦品种(系)的5个花药培养力性状进行了鉴定,并对5个性状进行了相关性分析。结果表明,74个基因型的愈伤组织诱导率、绿苗分化率、绿苗产率、白苗分化率及白苗产率变化范围分别为0~43.17%、0~139.29%、0~20.83%、0~63.33%、0~7.17%,各花药培养力性状在所研究基因型中差异明显,存在基因型依赖性,其中绿苗分化率基因型间差异最大。基因型愈伤组织诱导特性与绿苗、白苗的分化正相关,愈伤再生分化成绿苗或是白苗没有相关性。筛选出绿苗产率高于1.0%的基因型22个,其中,SPLM2、衡96851、石4185、邯6172、河农6425五个基因型农艺性状较好,绿苗产率依次为8.17%、5.44%、2.39%、2.00%、0.72%,可作为花培育种的骨干亲本。     

对马铃薯4x-2x,2x-4x及2x-2x杂种四倍体进行花药培养获得转育2n配子基因的双单倍体新个体

利用3个2n配子材料(2x)在马铃薯(S.tuberosum L.)中进行4x—2x,2x—4x和2x—2x的杂交,获得了4个四倍体杂种材料;然后对它们进行花药培养,共得到32个双单倍体植株。检查其中23个植株,有2株是具5％以上2n花粉粒的双单倍体,1株是重组了2n卵基因的双单倍体。由此证明花药培养的倍性操作技术是转育马铃薯2n配子性状给双单倍体的有效方法之一。     

Plant regeneration from tuber discs of potato (Solanum tuberosum L.) using 6-benzylaminopurine (BAP)

Summary Shoots, roots and callus were formed from tuber discs of potato, cultivar Désirée, when grown in vitro on the basal medium of Murashige & Skoog (1962) (MS) supplemented with 2,4-D and/or BAP. Callus was formed in MS medium with 1 mg l⁻¹ BAP plus 0.5 mg l⁻¹ 2,4-D, callus and roots were formed in MS with 1 mg l⁻¹ BAP plus more than 0.5 mg l⁻¹ 2,4-D and shoots were formed directly on tuber discs cultured on MS medium with 1 mg l⁻¹ BAP without the addition of 2,4-D. Nodules produced at the explant surface after the 4th week increased in size following subculture onto the same medium (MS+BAP alone), and 2 to 6 shoots developed from each nodule. After 9 weeks total time in culture, these shoots were excised and transferred as cuttings to MS medium without growth regulators, after which roots developed and plantlets were formed. A histological study of the explants at the sites of nodule formation indicated that the shoots developed from meristematic zones initiated within small outgrowths of tissue similar to those occuring in adventive organogenesis but the presence of shoot and root meristems associated with the same axis suggests the formation of somatic embryos.     

单倍体籼稻无性系微芽的离体调控

 以籼稻单倍体无性系微芽为材料,研究了激素及秋水仙碱等处理对籼稻单倍体微芽的诱导、分化、扩增及加倍的影响。2 mg/L的2,4-D可提高籼稻单倍体微芽的培养力。NAA对籼稻单倍体微芽的扩增有明显影响,0.5 mg/L的NAA可成倍扩增微芽,培养35 d后,其芽数比原始芽数增加了46.21倍,比不加NAA的对照增加了2.8倍,且单芽重仅0.079 mg。籼稻单倍体微芽扩增的适宜培养基为：N6附加MET 2.5 mg/L、NAA 0.5 mg/L、6-BA 2 mg/L。500 mg/L秋水仙碱溶液处理籼稻单倍体微芽48 h,其二倍体得率较高,绿苗率及二倍体率分别为42.9%和60.0%;秋水仙碱处理愈伤组织的效果不佳,虽然提高秋水仙碱处理浓度可提高二倍体率,最高可达100%,但由于绿苗分化率下降,使总的二倍体得苗率比不处理的对照低。     

芦荟库拉索(Aloe vera L)再生体系的建立

对芦荟(AloeveraL.)组织培养和再生体系进行了研究,初步获得芦荟的初代和继代培养基为MS 6-BA2.5mg/L NAA0.2mg/L 蔗糖30mg/L,生根培养基成分为1/2MS IBA0.2mg/L NAA0.1mg/L 蔗糖30mg/L,愈伤组织培养采用粗壮的芦荟基部的白色部分,诱导愈伤组织的培养基为B5 6-BA2.0mg/L 2,4-D2.0mg/L 蔗糖30mg/L,采用Vc5.0mg/L抑制愈伤组织诱导过程中褐变的产生。诱导愈伤组织和外植体直接出芽2种方式并存。试管苗移栽后存活率为100%。采用潮霉素为抗性筛选标记,选择浓度为20￣30mg/L。     

Effects of Various Phytohormones On Haploid Wheat Production in Wheat x Maize Crosses

Abstract

The Effects of Phytohormones On The Production of Haploid Wheat Were Examined in The intergeneric Crosses Between Japanese Wheat Cultivar (Triticum Aestivum Cv. Zenkojikomugi) and Maize (ZEA MAYS Cv. Pioneer P80 Lisa). The Detached Wheat Spikes Pollinated With Maize Were Cultured in A Solution Containing Sucrose and Sulfurous Acid Supplemented With 2,4―Dichlorophenoxyacetic Acid (2,4―D), indole-Acetic Acid, Naphthalene Acetic Acid, Kinetin Or 6-Benzylaminopurine At 0, 0.1, 1, 10, 100 Or 1000 Mg L-¹. Haploid Embryos Obtained Were Cultured On Agarose-Solidified B5 Medium. The Frequency of Plant Regeneration Was Significantly Affected Only By The Treatment With 100 Mg L-¹ 2,4―D. Thus, The Detached Spikes Were Cultured in The Medium Containing 2,4―D At 0, 25, 50, 75, 100, 125, 150, 175, 200 Or 400 Mg L-¹. The Treatments With 50 Mg L-¹ 2,4―D increased The Embryo Size, But The Treatments With Above 75 Mg L-¹ 2,4―D inhibited The Development of The Embryo. The Percentage of Florets Developing into Haploid Plants Was increased By The Treatment With 100 Mg L-¹ 2,4―D. Therefore, The Concentration of 2,4―D in THe Spikes Treated With 2,4―D At 50, 100 and 150 Mg L-¹ WERe Measured By Gas Chromatography. The Concentration of 2,4―D in THE Seed Was increased To 9.24 Ppm By The Treatment With 100 Mg L-¹ 2,4―D, A Further increase of 2,4―D Concentration in The Medium Having No Effect. On The Other Hand, The Concentration of 2,4―D in The Glumes and Rachis increased Up To 12.72 and 41.55 Ppm By The TreatmenT With 100 and 150 Mg L-¹ 2,4―D, Respectively. The Treatments With 2,4―D At A Concentration Higher Than 100 Mg L-¹ inhibited Embryo Development. The Present Results Suggested That 2,4―D At The Concentrations From 50 To 100 Mg L-¹ Would Be Optimum For Haploid Wheat Production Using Maize.     

An evaluation of callus induction and plant regeneration in twenty-five turf-type tall fescue (Festuca arundinacea Schreb.) cultivars

In order to investigate the genetic variation in tissue culture response and to find the cultivars with high regeneration ability for genetic transformation, twenty-five turf-type tall fescue ( Festuca arundinacea Schreb.) cultivars, including many elite ones released recently, were evaluated for their callus induction and plant regeneration responses. Callus induction was initiated from mature seeds on a Murashige and Skoog (MS) medium containing 9·0 mg l^–1 2,4-dichlorophenoxyacetic acid (2,4-D). Induced calli were subcultured on the same medium with 2·0 mg l^–1 2,4-D and then transferred to a MS medium supplemented with 2·5 mg l^–1 6-benzylaminopurine (BAP) for plant regeneration. Significant differences were observed among the twenty-five cultivars in both callus induction and plant regeneration ( P  < 0·001). Callus induction rate of viable seeds varied from 4·4% to 51·9%. Callus regeneration rates ranged from 16·7% to 58·8%. Overall regeneration rates (number of regenerated calli over number of cultured viable seeds) ranged from 1% to 22%. Approximately 94% of the regenerants were green plantlets.     

节节麦幼胚再生体系的建立

为建立高效的节节麦幼胚再生体系,以节节麦幼胚为外植体,通过正交设计,探索基本培养基、2,4-D、碳源、KT等因素对幼胚愈伤组织诱导、分化及植株再生效果的影响。结果表明,4种因素中,基本培养基对节节麦幼胚愈伤组织诱导的影响最显著(P0.05),2,4-D浓度对节节麦幼胚愈伤组织诱导也有显著影响(P0.05),添加3.0mg·L~(-1) 2,4-D的培养基诱导出的愈伤组织质量高,淡黄色,表面呈不规则颗粒状,质地致密,再生频率可以达到17.62%。KT浓度对节节麦愈伤分化影响最显著,基本培养基、2,4-D和碳源对节节麦幼胚愈伤分化均无显著影响。不同碳源对节节麦幼胚愈伤组织诱导和分化的影响均不显著,为节约成本可直接选用30g·L~(-1)蔗糖作为碳源。节节麦幼胚组织培养的最佳组合是:愈伤诱导培养基为MS培养基+3mg·L~(-1) 2,4-D+15g·L~(-1)蔗糖+15g·L~(-1)甘露醇,愈伤分化培养基为MS培养基+15g·L~(-1)蔗糖+15g·L~(~(-1))甘露醇+1.0mg·L~(-1) KT。     

Effect of temperature on the starch content of potato callus tissue

Starch content was determined in Lemhi Russet callus grown on MS medium for 2 and 4 weeks at 25, 15, and 5°C with 50 μM 2,4-D or 10 μM picloram as the only plant growth regulator. A treatment of 5°C for 4 weeks followed by 25°C for 2 and 4 weeks was included to simulate reconditioning. Callus starch content ranged from 0.01 to 0.08% on a wet weight basis and 0.5 to 4.0% on a dry weight basis. Starch content was highest after 2 weeks at 25 and 15°C and decreased with time in a similar manner at both 25 and 15°C. Starch content decreased at 5°C, then increased significantly during simulated reconditioning at 25°C. Starch content was higher in callus grown with 50 μM 2,4-D than in callus grown with 10 μM picloram. After staining the callus with I₂-KI solution, starch granule formation was observed in the callus grown with 50 μM 2,4-D, while little granule formation was observed in the callus grown with 10 μM picloram.     

玉米愈伤组织再生体系的研究

用玉米胚性愈伤组织作为转化受体时,不同基因型在相同2,4-D浓度的培养基上诱导,其诱导率有较大差别,齐319最高;胚龄对诱导率的影响也较大,10d左右最易诱导,过大或过小均不易诱导胚性愈伤;培养基中2,4-D的浓度对玉米愈伤组织的诱导率有极大影响,不同基因型的最大胚性愈伤组织诱导率所需的2,4-D浓度也不相同,一般为0.5～1.0mg/L。愈伤组织分化时不同基因型所需最适6-BA浓度不同,齐319、515的愈伤组织最适合分化成苗的6-BA浓度为1mg/L,鲁原341愈伤组织最适6-BA浓度为0.5mg/L。     

快中子辐照对花生种子胚小叶植株再生的影响

以花生品种花育22号的成熟饱满种子为试材,采用14Mev不同剂量的快中子(0、9.7、14和18Gy) 进行辐照处理。处理后的种子经表面杀菌后,取胚小叶作为外植体先后在添加2,4-D 和BAP 的培养基上进行培养,诱导体胚形成及其萌发和植株再生。结果表明,体胚诱导率和植株再生率因辐照剂量的不同表现出明显的差异,随辐照剂量的增加,外植体形成体胚的频率及再生植株的频率明显降低。推断快中子辐照花育22号的适宜剂量为9.7~14.0Gy。再生小苗经无菌嫁接驯化后移栽田间,得到了成熟种子。           

大豆花药培养几个问题的研究

本文研究了大豆花药培养中的培养基、基因型、激素配比、糖分种类及浓度、取材时期、预处理温度、接种方式、有机添加物等因素对愈伤组织诱导频率的影响。认为大豆花药在培养基上的脱分化启动具有群体效应，合适的取材时期是单核中晚期。高浓度蔗糖能抑制体细胞愈伤组织的产生，而愈伤组织的分化则需要较低的蔗糖浓度。愈伤组织在Ｂ＿５＋０．５ｍｇ／１ＮＡＡ＋１．０ｍｇ／１ＫＴ＋１％蔗糖和改良ＭＳ＋０．１ｍｇ／１ＩＢＡ＋０．１ｍｇ／１ＧＡ＿３＋０．４ｍｇ／１ＮＡＡ＋０．５ｍｇ／１ＢＡ＋０．５ｍｇ／１ＫＴ＋０．５ｍｇ／１ＺＴ＋０．５ｍｇ／ｌ生物素＋２％蔗糖等培养基上分化出了芽，在改良ＭＳ＋０．５ｍｐ／１ＩＢＡ＋０．５ｍｇ／１ＢＡ＋０．５ｍｇ／１ＫＴ＋０．５ｍｇ／１ＺＴ＋５％蔗糖＋１％麦芽糖等培养基上产生了胚状体。