基因型和激素浓度对大豆植株再生的影响

本文采用8个大豆基因型,比较了NAA 激素浓度对大豆下胚轴植株再生的影响。结果表明：所有的基因型在合适的培养条件下, 均能形成愈伤组织,但不同基因型由愈伤组织诱导植株再生频率有很大差异。供试的8个品种中,在NAA浓度为0.1～0.5 mg／L的培养基上,汾豆33 号表现出较高的再生率,达50%;晋豆19号次之,为42%;而晋旱125则较低,仅19%。较合适的培养条件是：下胚轴在B5+NAA 0.3 mg／L+KT 1 mg／L+3%蔗糖,可高频率地诱导出愈伤组织并直接分化成苗。     

罗布麻子叶和下胚轴再生植株的培养

本文报道了用罗布麻（Apocynum venetam)幼苗的子叶和下胚轴切段诱导出愈伤组织和再生植株。结果表明,愈伤组织的诱导在附加0.5mg/L6－BA和0.1-1mg/L NAA的MS培养基上为最好。在附加0.5mg/L NAA的MS培养基上培养愈伤组织能促进芽的分化,当NAA浓度增加到1mg/L时则能抑制芽的分化。随后在附加0.4mg/L IBA的MS培养基上诱导生根,获得完整再生植株。     

甘蓝型油菜子叶和下胚轴再生植株无性系建立

以甘蓝型油菜(Brassica napus L.)豫油2号和6257的子叶和下胚轴为材料,在不同激素配比的MS培养基上诱导出了愈伤组织。将经过继代的部分愈伤转入分化培养基,分化结果表明:除基因型、外植体和分化培养基的激素配比对分化率有影响外,诱导愈伤培养基的激素配比对分化率也至关重要。豫油2号的子叶和下胚轴在最适诱导培养基(ZT 1+NAA0.5+2,4-D 0.2 mg/L)和最适分化培养基(ZT4+IAA 0.2 mg/L)组合中的愈伤分化率分别为12.5%和75%;6257的子叶和下胚轴在其最适诱导培养基(KT 2+NAA1+2,4-D 0.2 mg/L)和最适分化培养基(6-BA 4+IAA 0.02 mg/L)组合中的愈伤分化率分别为50%和37.5%。将其最适诱导培养基中的愈伤组织继代达8个月以上,建立了不同继代愈伤的再生植株无性系。     

六个海岛棉品系体细胞胚胎发生及再生体系的研究

以海岛棉DJ-1(373)、天长2号(376)、5917(380)、阿长-599(381)、塔07-152(383)、S0717(408)6个品系的下胚轴为外植体,通过双因素随机区组设计的方法筛选出的两组最佳激素组合,诱导出愈伤组织,并成功获得再生植株。研究结果表明,0.1 mg/L 2,4-D+0.1 mg/L KT为海岛棉380、383和381愈伤组织最佳诱导条件,0.3 mg/L NAA+0.1 mg/L KT为海岛棉373、376和408愈伤组织最佳诱导条件;诱导出的愈伤组织接种于KNO3加倍的培养基可产生胚性愈伤组织;将其接种于含有0.1mg/L KT和0.05 mg/L IBA的培养基中可促进体细胞胚胎的发生和成熟,利用畸形苗的培育可缩短再生体系的周期,6个品种在6-8个月内均能获得再生植株。     

党参的离体培养及植株再生的研究

在附加激素的MS培养基上,培养党参下胚轴和无菌芽切段,诱导产生愈伤组织并且再生植株。经过两年多(15个世代)的继代培养,建立了党参体细胞无性系。实验结果表明:(1)培养基MS 0.4mg/L 2,4-D 0.8mg/L Kt 2.0mg/L IAA对愈伤组织诱导及继代培养,MS 0.2mg/L 6-BA诱导外植体产生丛芽和愈伤组织再分化,MS 0.5mg/L NAA 0.2mg/L 6-BA及MS 0.2mg/L NAA诱导生根效果最好。(2)愈伤组织再分化经过胚状体途径。     

苦荞胚性愈伤组织诱导与植株再生研究

以苦荞子叶和下胚轴为外植体,进行了不同浓度激素组合的MS和SH固体培养基对胚性愈伤组织诱导及植株再生的研究。结果发现,MS培养基比SH培养基更有利于胚性愈伤组织诱导;2,4-D是诱导愈伤组织的有效激素,KT能有效促进胚状体的形成;下胚轴和子叶都能有效诱导出胚性愈伤组织和再生植株。下胚轴在MS 1.5mg·L-12,4-D 1.5mg·L-1BA培养基,子叶在MS 2mg·L-12,4-D 0.5~1.5mg·L-1BA上能高效诱导出愈伤组织;愈伤组织在MS 2mg·L-12,4-D 0.1mg·L-1KT培养基中继代,能有效诱导胚性愈伤组织;来自下胚轴的胚性愈伤组织在1/2MS 2.0mg·L-1BA 0.5mg·L-1KT 0.1mg·L-1NAA培养基上能够高频再生出芽,来自子叶的胚性愈伤组织在1/2MS 1.0mg·L-1BA 0.1mg·L-1KT 0.1mg·L-1NAA培养基上芽诱导率较高;MS 1mg·L-1NAA是适宜的再生苗生根培养基。     

怀山药微型块茎愈伤组织的诱导形成及高频率再生

对怀山药微型块茎愈伤组织的诱导及高频率再生进行了研究。结果表明 :(1)光下诱导铁棍山药脱分化形成愈伤组织 ,6 -BA2 mg/L NAA2 mg/L 为最佳激素组合。KT2 mg/L 2 ,4 - D2 mg/L 有利于铁棍山药愈伤组织的增殖。附加 2 mg/L NAA能缩短 4 7号山药愈伤组织的诱导时间。KT2 mg/L NAA2 m g/L 有利于 4 7号山药愈伤组织增殖 ;(2 )不同光照条件对愈伤组织的诱导和增殖影响不同。光照是缩短 4 7号山药愈伤组织诱导时间的另一因素。暗培养有利于愈伤组织的增殖 ,对 4 7号山药来说 ,暗培养下诱导率也较高 ;(3)基因型不同 ,愈伤组织类型不同 ,诱导率和不定芽分化率也有差异 ,4 7号山药高于铁棍山药 ;(4 )KT对 4 7号山药愈伤组织分化形成不定芽起主要作用 ,2 ,4 - D2 mg/L KT2 mg/L 为最佳激素组合 ;(5 )光培养有利于不定芽的分化     

6-BA、NAA和2,4-D不同配比对荠菜愈伤组织诱导、生长及植株再生的影响

以野生荠菜[Capsella bursa-pastoris （L.） Medic]无菌苗为试材,选取下胚轴、子叶、真叶和叶柄作为外植体,研究了不同外植体的出愈情况,不同植物生长调节剂及配比对愈伤组织诱导、继代及植株再生的影响。结果表明：（1）下胚轴作为外植体出愈情况最好,继代后生长快;（2）下胚轴愈伤组织的最适植株再生培养基为MS＋2～3mg·L-16-BA＋0.2~0.6mg·L-1NAA;（3）愈伤组织培养阶段的2,4-D浓度对其植株再生能力有影响。     

沙打旺原生质体培养再生植株

用1%半纤维素酶,0.4%纤维素酶,0.1%果胶离析酶,CPW9M酶液分离沙打旺无菌苗下胚轴和子叶原生质体。K8P原生质体培养基悬滴培养。下胚轴原生质体形成小细胞团后用琼脂糖包埋培养,形成小块愈伤组织后转入增殖培养基M1、M2(改良MS培养基)上形成大块愈伤组织。经过两次诱导分化,在分化培养基M3(MS 0.7mg/L BA 0.2mg/L NAA),M4(MS 0.5mg/L BA 0.5mg/L KT 0.5mg/L ZT 0.2mg/L NAA)和M6(MS 3mg/L ZT 0.2mg/L IAA)上分化出苗,再生植株。由子叶分离的原生质体未能形成愈伤组织。     

稀有植物裸果木的组织培养及植株再生

对稀有植物裸果木进行组织培养研究,在MS培养基上裸果木的下胚轴脱分化形成愈伤组织,并进一步分化形成再生植株。激素种类及其浓度是器官脱分化与植株再生的决定因素。诱导下胚轴形成愈伤组织的最适培养基为MS 1mg／L 6-BA 0．5mg／L NAA;芽分化诱导的最适培养基为MS 1 mg／L6-BA;生根的最适培养基为不含任何激素的1／2MS培养基。     

An efficient in vitro regeneration system of fieldpea (Pisum sativum L.) via. shoot organogenesis

In-vitro regeneration in fieldpea was achieved from immature embryonic axes and cotyledonary node explants of six genotypes on modified MS media supplemented with different concentration of plant growth regulators, 6-Benzylamino purine (BAP) and Naphthalene acetic acid (NAA). The best regeneration response, leading to multiple shoot formation efficiency (22.34 shoots/explant) was observed in the medium supplemented with 1.0 mg/L BAP and 0.2 mg/L NAA and best frequency (67.55?±?4.74) was achieved on medium containing 2.0 mg/L BAP and 0.4 mg/L NAA. The shoots were subcultured on a medium supplemented with a combination of 1.0 mg/L GA₃, 2.0 mg/L BAP and 0.4 mg/L NAA, which resulted in elongation of 85 % of shoots. Rooting attempted from the elongated shoots, on half strength MS medium and supplemented with three different auxins IBA, IAA and NAA separately, exhibited similar results. Alternatively, micro-grafting of in vitro regenerated shoots onto pre-germinated root stocks raised in green house facility was attempted with high success rate (75 %). The grafted plants could be successfully hardened, fertigated with Hoagland solution and distilled water in a ratio of (1:10) for acclimatization and further development. All the genotypes tested, produced multiple shoots that could be established to mature fertile plant, hence, the medium combinations used were found to be genotype neutral.     

一种快速、高效花生植株再生体系的建立

快速、高效、重复性好的植株再生体系是转基因育种的基础;本研究以14份不同花生品种的胚小叶为外植体,利用不同激素浓度、组合和不同花生基因型筛选最佳芽诱导培养基、伸长培养基和高效再生基因型。结果表明最佳丛生芽诱导培养基为MSB;＋0．2mg·L-1NAA＋6mg·L-1 6-BA,诱导率为89．50％;最佳伸长培养基为MSB5＋0．2mg.L-1 NAA＋3mg’L-1 6-BA和MSB;＋O．2mg·L～NAA＋4mg·L-1 6-BA＋2mg·L～GA,交替培养,每个丛生芽伸长数达到7．24,时间缩短至3-4周。不同品种再生率的变幅在25．51％～93．01％,大于80％的品种有‘麻油1-1’、‘弗落蔓生’、‘濮花23号’、‘海花1号’。利用‘弗落蔓生’在15周内得到了生根组培苗。     

吉粳88高效再生体系的建立

以吉粳88为材料,通过对不同培养基和激素的筛选,确定最佳的再生体系。结果表明:NMB培养基为吉粳88最适的愈伤组织诱导培养基。通过L₉(3³)正交实验研究最适激素浓度配比,筛选出吉粳88愈伤组织诱导的最佳激素配比为2.0 mg/L 2,4-D+0.2 mg/L 6-BA+0.10 mg/L NAA。在添加2.0 mg/L 6-BA+0.5 mg/L NAA的NMB基本培养基条件下,吉粳88的分化率最高,成培苗率也最高。     

彩色马蹄莲的组织培养

以彩色马蹄莲芽眼为外植体,在添加不同激素配比的MS培养基上进行培养.结果表明:(1)较适宜诱导愈伤组织的激素组合是1.5 mg/L Zt+0.1 mg/L NAA、3.0 mg/L 6-BA+0.1 mg/L NAA或0.5 mg/L 2,4-D+0.1 mg/L NAA,出愈率分别为94.12%、100%和86.67%;较适宜诱导不定芽的激素组合是2.0 mg/L 6-BA+0.1 mg/L NAA,芽丛诱导率可达87.3%;芽增殖较适宜的激素组合是2.0 mg/L 6-BA+0.1 mg/L NAA或2.0 mg/L KT+1.0 mg/L NAA,而且在继代培养中,交替使用这两种激素组合,繁殖系数可达4倍以上;生根培养基以1/2 MS+0.3 mg/L NAA+0.2 mg/L IAA较为适宜,生根率达95%以上.(2)彩色马蹄莲小块茎形成的主要影响因素是培养基中的蔗糖浓度,最适宜蔗糖浓度为10%;10 mg/L多效唑和8 mg/L NAA对小块茎的形成有明显的促进作用,而分裂素则表现出较强的抑制作用.     

播娘蒿下胚轴高频率再生植株因素的研究

以播娘蒿（Descurainia sophia)下胚轴为外植体,研究影响其分化的若干因素。结果表明：用附加2,4－D的培养基,对下胚轴切段预培养5d可提高分化频率;1.7mg/L AgNO3对分化出芽有很大促进作用,最佳激素组合为2.0mg/L 6-BA 0.2mg/L NAA;黄化苗的下胚轴比绿苗的下胚轴对激素反应更敏感,最高分化频率可达86.05%。再生芽转至1／2MS 0.5mg/L NAA 0.5mg/L IBA的培养基中,生根率100％。     

马铃薯离体再生及人工种子研究

马铃薯３个品种虎头,克４和Ｆａｖｏｒｉｔａ的茎叶外植体在ＭＳ＋１ｍｇ／ＬＮＡＡ＋１ｍｇ／ＬＢＡ培养基上形成愈伤组织。在ＭＳ＋０２ｍｇ／ＬＮＡＡ＋１ｍｇ／ＬＢＡ培养基上,愈伤组织分化产生不定芽。在ＭＳ＋００５ｍｇ／ＬＮＡＡ培养基上,不定芽生根形成再生完整植株。０２～０３ｃｍ大小的不定芽反复继代可持续增殖。有３个以上叶片的不定芽在ＭＳ＋５ｍｇ／ＬＢＡ＋００５ｍｇ／ＬＩＢＡ培养基上和黑暗条件下,在侧芽或顶芽部位形成微型薯。用４％海藻酸钠和２％氯化钙溶液包裹０２～０３ｃｍ大小的不定芽或直径为０２～０３ｃｍ的微型薯制成微芽人工种子和微薯人工种子。在４℃下贮存２个月后,微芽人工种子和微薯人工种子在有菌腐殖土壤中播种２１ｄ的萌发率分别是１５７％和９６２％。     

Influence of growth regulators on plant regeneration from epicotyl and hypocotyl cultures of two groundnut (Arachis hypogaea L.) cultivars

This study was designed to evaluate the effect of phytohormones on plant regeneration from epicotyl and hypocotyl explants of two groundnut (Arachis hypogaea) cultivars. Explants cultured on media with auxins and in combination with cytokinin produced high frequency of callus. After four weeks, callus from these cultures was transferred to medium with cytokinin and reduced auxin, shoot buds regenerated from the cultures. A high rate of shoot bud regeneration was observed on medium supplemented with 2.0 mg/L BAP and 0.5 mg/L NAA. Among the different auxins tested, NAA was found to be most effective, producing the highest frequency of shoot buds per responding cultures. Of the two explants tested, epicotyl was found to be best for high frequency shoot bud regeneration. Multiple shoots arose on MS medium supplemented with BAP or kinetin (1.0–5.0 mg/L) plus IBA (1.0 mg/L), with maximum production occurring at 5.0 mg/L. The elongated shoots developed rootsin vitro upon transfer to MS medium supplemented with NAA or IBA (0.5–2.0 mg/L) and kinetin (0.5 mg/L) for 15 days.In vitro produced plantlets, were transferred to soil and placed in a glasshouse developed successfully, matured, and set seeds.     

佳禾早占成熟种胚高效率组织培养方法的研究（简报）

In order to improve the embryogenic callus induction rate and the regeneration rate of JiaHe-ZaoZhan rice, the influence of different factors were investigated, media with different hormones were used. Induction medium was supplemented with 1.5 mg/L 2,4-D + 3 mg/L NAA + 0.1 mg/L KT + 1 mg/L phytic acid + 20 mg/L PAA. Embryogenic call were treated under the condition of 25 degrees C before transferring to regeneration medium, the regeneration medium contained 0.5 mg/L 6-BA + 3 mg/L NAA + 0.5 mg/L KT + 1 mg/L phytic acid. The experiment results indicated that the hormone treatments had certain effects on the callus induction. Under the optimal medium, culture condition and the hormone treatments, the embryogenic callus induction could reach over 95%, and dry treatment of embryogenic callus had been found to increase the frequency of plant regeneration, significantly the plant regeneration rate could reach over 80%. Transplanted into pots, the young plants grew well. Then a experimental system with stability and high regenerating efficiency has been established for the mature seeds of rice (JiaHe-ZaoZhan).     

Combination of 6-Benzylaminopurine and Thidiazuron Promotes Highly Efficient Shoot Regeneration from Cotyledonary Node of Mature Peanut (<i>Arachis hypogaea</i> L.) Cultivars

Efficient in vitro plantlet regeneration is an important step to successfully transform genes for the improvement of agronomic traits. A combination of 6-benzylaminopurine (BAP) and thidiazuron (TDZ) plant growth regulators was applied to evaluate shoot regeneration capacity whereas α-naphthalene acetic acid (NAA) combination with 6-benzylaminopurine (BAP), and 2, 4-dichlorophenoxyacetic acid (2, 4-D) with 6-benzylaminopurine were tested to optimize root induction for two peanut cultivars. The result showed combination (BAP with TDZ) was found to be effective in promoting shoot. The highest shoot regeneration frequency (93%) was obtained on a medium supplemented with 4 mg/L BAP and 0.5 mg/L TDZ while an average regeneration frequency (87%) was achieved in a medium containing combinations of 2 mg/L BAP with 1 mg/L TDZ. The shooting rate increased for both cultivars as the concentrations of BAP increased and TDZ decreased. The highest rooting rate (93%) was obtained on a medium supplemented with 3.5 mg/L NAA with 2.5 mg/L BAP for both cultivars. The rooting rate increased as the concentration of auxin to cytokinin ratio increased. The maximum rooting rate (83%) was obtained on MS medium supplemented with 0.3 mg/L 2, 4-D with 0.2 mg/L BAP for the cultivar N3. The result indicated that BAP with NAA was much better than BAP with 2, 4-D in rooting rate. Thus, the protocol developed was genotype independent and effective for peanut tissue culture.