藏红花瓣状体的诱导与花柱-柱头状物的定向分化

藏红花(Crocus sativus L.)又名番红花、西红花;藏药中称为苟日苟木.它是鸢尾科草本植物,原产于欧洲、地中海地区.我国长期以来把它作为珍贵的中藏药.其药用部位是柱头,有效成分主要是藏红花素、藏红花酸、藏红花醛和藏红花苦素.丁葆祖等于1979年首次从藏红花的球茎获得完整的植株[1].Sano等人1987年在离体条件下,诱导花柱-柱头状物再生获得成功[2],此后国内外不断有相关报道[3～7].但试验大多集中在由外植体直接或由愈伤组织间接诱导柱头状物,且花柱-柱头状物的频率低和数量少.我们试图先由藏红花花瓣诱导瓣状体,再定向分化花柱-柱头状物,来提高花柱-柱头状物的诱导频率和数量.从而为以后的大规模生产奠定基础.     

番红花雄蕊柱头状物的离体再生

以番红花（Crocus sativus L.)雄蕊为材料诱导培养出花柱柱状结构,诱导率可达30％,起源于花丝基部。影响雄蕊柱头状物诱导的主要因素为外植体的发育期和生长素NAA的使用浓度。幼嫩浅黄色雄蕊适于诱导柱头状物。温度和光照在不同激素水平下对雄蕊柱头状物诱导的影响不同。紫外检测表明,由雄蕊诱导出的柱头状物含有番红花甙、番红花醛和番红花苦甙。其含量明显高于由药柱诱导出的柱头状结构。在诱导花魔王、子房、花瓣的柱头状物的过程中,观察到成药逆转现象。     

彩色大白菜子叶的不定芽再生

以彩色大白菜子叶为外植体,研究不同激素配比和AgNO3对不定芽再生的影响。结果表明:单独附加细胞分裂素(6-BA或TDZ)的MS培养基,不能诱导子叶不定芽分化;而同时附加生长素(NAA)和细胞分裂素(6-BA或TDZ),不定芽的再生频率提高,最高为15%;AgNO3与细胞分裂素及生长素配合使用,能大幅度提高子叶不定芽的再生频率,提高率最高达42.5%。与6-BA相比,TDZ对不定芽再生的效果更好。当TDZ浓度为0.05mg/L、NAA为0.3mg/L、AgNO3为8mg/L时,产生丛状芽数目最多,再生率最高,达50%。     

樱桃砧木Colt离体叶片再生

以樱桃砧木Colt试管苗的叶片为外植体 ,通过先诱导愈伤组织分化不定芽以及叶片直接分化不定芽两种途径诱导再生。结果表明 :在MS附加NAA 1 0mg/L、KT3 0mg/L、ZT0 2 5mg/L培养基中 ,愈伤诱导率可达 1 0 0 % ;诱导的愈伤在MS附加NAA 0 2mg/L、IAA0 5mg/L、6 BA 0 5mg/L、KT 1 0mg/L、GA 0 5mg/L培养基中 ,不定芽分化率为 2 1 3% ;在MS附加 6 BA 6 0mg/L、NAA 1 0mg/L、GA 0 5mg/L中 ,叶片 -叶柄不定芽诱导率可达 48 3%。     

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。     

花叶千年木愈伤组织直接再生花序

以花叶千年木(Dracaena fragrans cv. Massangeana Hort.)的花被筒、花序分枝轴和花序轴为外植体成功地诱导了花序的直接再生. 3种外植体首先在MS附加1.0 mg/L 6-BA和0.5-0.8mg/L 2,4-D的培养基上诱导形成愈伤组织,然后转移到MS附加0.5 mg/L 6-BA和0.005～0.5 mg/L 2,4-D的培养基上分别诱导了花序的直接再生.观察了愈伤组织形成和花序分化的形态学过程.     

花叶千年木愈伤组织直接再生花序

以花叶千年木（Dracaena fragrans cv.Massangeana Hort.)的花被筒，花序分枝轴和花序轴为外植体成功地诱导了花序的直接再生，3种外植体首先在MS附加1.0mg/L6-BA和0.5-0.8mg/L2,4-D的培养基上诱导形成愈伤组织，然后转移到MS附加0.5mg/L6-BA和0.005-0.5mg/L,2,4-D的培养基上分别诱导了花序的直接再生，观察了愈伤组织形成和花序分化的形态学过程。     

甘蓝型油菜高效离体再生体系的建立

以甘蓝型油菜(Brassica napusL.)HC8为材料,从无菌苗苗龄、预培养基激素浓度、预培养天数、6-BA及NAA的浓度等方面对影响油菜组织培养的因素进行了分析研究,建立了甘蓝型油菜品系HC8的离体再生技术体系。结果表明,该油菜组织培养的最佳苗龄为5 d;最佳预培养时间为5 d,最佳2,4-D浓度为1.0 mg/L;子叶最佳诱导培养基为MS+2.0 mg/L 6-BA+0.05 mg/LNAA+3.5 mg/L AgNO3或MS+3.0 mg/L 6-BA+0.1 mg/L NAA+3.5 mg/L AgNO3,该条件下子叶愈伤组织诱导率最高可达100%,再生频率及分化频率分别可达88.0%和108.33%;下胚轴最佳诱导培养基为MS+4.0 mg/L 6-BA+0.05 mg/L NAA+3.5 mg/L AgNO3,子叶愈伤组织诱导率最高可达95.24%,再生频率及分化频率分别可达81.82%和104.55%;最佳生根培养基为MS+0.5 mg/L NAA,生根率最高为90.0%。     

甜菜叶柄高效直接再生体系的建立

通过预试验,从4中不同基因型甜菜中选取KWS-9103作为试验材料,建立其叶柄高效直接再生体系。通过种植无菌苗、预培养、诱导分化培养,获得了再生植株,建立了甜菜快速繁殖体系。结果表明,甜菜种子经过消毒处理后培养,取幼苗在MS附加6-BA 0.5 mg/L和NAA 0.05 mg/L的培养基中预培养,再取叶柄作为外植体转入诱导培养基MS附加6-BA0.5 mg/L和NAA0.05 mg/L中诱导不定芽,不定芽诱导率为50%以上。在MS附加IBA2.0 mg/L生根培养基中生根,诱导生根率在90?以上,移栽成活率高达95%。     

水稻单倍体幼穗切块的离体培养

长0.5—4厘米的水稻单倍体幼穗,剪切成1毫米左右的碎块,培养在附加2.4-D 1mg/l的N_6培养基上,切块容易被诱导生成愈伤组织。愈伤组织转移到附加NAA0.5-KT 2mg/l的培养基上培养,约30%分化出苗。由单个幼穗的全部切块平均可得30丛绿苗。幼穗切块培养在附加NAA 1 KT 2mg/l的培养基上,切块上颖花不孕稃片与内外稃之间的表皮和皮层细胞被诱导分裂并形成芽和根。通过这种直接出苗方式,每个幼穗的全部切块平均出苗12丛。由上述二种途径再生的小苗中,白化苗均低于2%。98％的再生植株仍为单倍体。用含有2%二甲基亚砜和200mg/l秋水仙碱的培养液浸泡愈伤组24小时(26℃),再生的植株中,能育二倍体植株达50%。     

Study on in Vitro Regeneration of Style-Stigma-Like Structure in Crocus sativus L.

Effects of age and variety of explant as well as of exogenous hormones on regeneration of style-stigma-like structure in vitro were observed in saffron. (1) Explants excised from stalk, perianth, anther, ovary, style and stigma showed different reactions in vitro, only young perianth and style could regenerate the style-stigma-like structures; (2) Age of the perianth explant had obvious effect on induction frequency, and the explants excised from an inflorescence of 24mm in length showed the highest frequency (37.5%) and (3) Experiments on exogenous hormones made clear that supplement with K 5mg/L and NAA 4mg/L was advantageous to regeneration of style-stigma-like structure. Induction frequency reached 35.3%. Supplement with 6-BAP 7mg/L and NAA 9mg/L could promote the formation of stigma like structures.     

In Vitro Induction of Regeneration of Fruit-Like Structure in Lycopersicon esculentum Mill.

It is possible to induce in vitro regeneration of fruit-like structures from tissue pieces excised from young pistil and young fruit of Lycopersicon esculenturn Mill. These fruit-like structures could be cultured in vitro to maturation, meanwhile they became red coloured. Under dissection it was observed that the fruit-like structure consists of fruit flesh and fruit coats without seeds and plancentae. Tests of exogenous hormones and explant ages revealed: 1. Supplement with exogenous cytokinins alone could induce the explants excised from the pistils to regenerate the fruit-like structures, and the highest induced frequency (50%) was obtained when 0. 5 mg/L zeatin was supplemented. Exogenous auxin seems unnecessary. 2. When tissues of young fruit were used as explants, all explants excised from 4—12 mm (diameter) young fruits could be induced to differentiate the fruit-like structures on medium supplemented with exogenous hormones. However, the hightest frequency of induction (62.5%) was obtained only in tissues of explants excised from 8 mm (diameter) sized fruit and cultured on medium supplemented with 2 mg/L of 6-BAP and 1 mg/L of NAA where regermination of the fruit-like structures took place. In order to investigate the expression of cell totipotency arisen in the regeneration of the fruit-like structure a concept of partial expression of plant cell totipotency was proposed and discussed.     

藏红花瓣状体的诱导与花柱-柱头状物的定向分化

藏红花 (ＣｒｏｃｕｓｓａｔｉｖｕｓＬ .)又名番红花、西红花 ;藏药中称为苟日苟木。它是鸢尾科草本植物 ,原产于欧洲、地中海地区。我国长期以来把它作为珍贵的中藏药。其药用部位是柱头 ,有效成分主要是藏红花素、藏红花酸、藏红花醛和藏红花苦素。丁葆祖等于 1979年首次从藏红花的球茎获得完整的植株[1 ] 。Ｓａｎｏ等人 1987年在离体条件下 ,诱导花柱 柱头状物再生获得成功[2 ] ,此后国内外不断有相关报道[3～ 7] 。但试验大多集中在由外植体直接或由愈伤组织间接诱导柱头状物 ,且花柱 柱头状物的频率低和数量少。我们试图先由藏红花…     

风信子花被外植体年龄对花器官分化的影响

离体培养风信子(Hyacinthus orientalis L．)不同年龄的花被外植体诱导花器官直接再生的实验表明;1．在MS附加6-BA 2 mg／L,2,4-D 0.1 mg／L的培养基上,年龄V的外植体大量衰老,基本丧失器官再生能力,处于年龄段Ⅱ～Ⅳ的外植体可发生玻璃化反应。2．玻璃化反应的外植体转移至MS附加6-BA 0.2 mg／L、NAA 0.005 mg／L的培养基上继续培养30 d后可再生正常的花被片,表明降低培养基中外源激素浓度能够阻止玻璃化反应继续发生。3．在MS附加6-BA 2 mg／L．2,4-D 0.1 mg／L的培养基上,外植体形态学下部可再生雌蕊状早期结构。平均每块外植体分化雌蕊状早期结构数以年龄Ⅲ的外植体最多。     

In Vitro Regeneration of Style-stigma-like Structure from Stamens of Crocus sativus

Style-stigma-like structures were regenerated from stamens of Crocus sativus L. The age of the stamen explant has an obvious effect on the induction rate. Auxin NAA has larger effect on the induction of filament style-stigma-like structure. Auxin NAA of higher concentration can lead to higher induction rate. Temperature and light have different effects on the induction of style-stigma-like structure from anther's filament of C． sativus with exogenous hormones at different levels. Ultraviolet tests show that style-stigma-like structure from anther's filament of C． sativus contains crocin, safranal and picrocrocin, contents of which are obviously more than those contained in the style-stigma-like from style. Floral reversion was observed in the induction of style-stigma-like structure from petals, ovaries and styles.     

Organogenesis and Plantlet Regeneration in Vitro of Populus euphratica

The research of organogenesis and in vitro plantlet regeneration of Populus euphratica Oliver was carried out using the tender shoots from mature tree as initial explants and MS medium as the basic medium. The effects of plant growth regulators (PGR) on the regeneration were compared. The results showed that the concentration of PGR was not strictly required for the organogenesis of the excised organs and callus, but the ratio of BA to NAA was important. Calli could be induced from the excised leaves and stems cultured on the medium with 0.5 mg/L BA and 0.5 mg/L NAA. The embryonic callus could be multiplied in dark on the medium supplemented with 0.25 mg/L BA and 0.5 mg/L NAA. For the adventitious bud regeneration of the leaf and callus, supplement with 0.5 mg/L BA and 0.1 mg/L NAA was appropriate, giving a regeneration frequency of 82.9% and 100%, respectively. The suitable level of BA and NAA for the excised stem's was 0.1 mg/L and 0.01 mg/L respectively, yielding a regeneration frequency of 83 %. Rooting occurred on the MS medium with half strength of macronutrient and addition of 0.015 mg/L NAA, and the rooting rate could reach up to 86.2%. The techniques of somatic cell cloning of P. euphratica was established in vitro. The problems of deterioration of the subcultured shoots were also discussed.     

外源激素在诱导风信子花被外植体不同部位细胞再生花芽中的作用

外源激素诱导风信子（Ｈｙａｃｉｎｔｈｕｓ ｏｒｉｅｎｔａｌｉｓＬ．）同一发育时期花被外植体不同部位细胞再生花芽的实验表明∶１． 诱导花被外植体细胞再生花芽,外源激素是必需的;２． 仅有细胞分裂素就可以诱导花芽再生,生长素并不是必需的;３． 花被外植体上的不同部位的细胞再生花芽时,需要不同浓度的外源激素． 单独加６－ＢＡＰ或玉米素２ ｍ ｇ／Ｌ可以诱导花被下部的细胞再生花芽;６－ＢＡＰ或玉米素２ ｍ ｇ／Ｌ和２,４－Ｄ ０．１ ｍ ｇ／Ｌ的组合有利于花被中部的细胞再生花芽;６－ＢＡＰ或玉米素２ ｍ ｇ／Ｌ和２,４－Ｄ １．０ ｍ ｇ／Ｌ的组合能促进花被上部的细胞分化花芽     

胡杨离体器官发生及试管无性系的建立

研究了离体条件下胡杨（Ｐｏｐｕｌｕｓ　ｅｕｐｈｒａｔｉｃａ　Ｏｌｉｖｅｒ）茎段、叶片及愈伤组织的器官发生和植株再生技术。离体培养以ＭＳ为基本培养基并附加４０ｍｇ／Ｌ腺嘌呤和５００ｍｇ／Ｌ水解乳蛋白。离体叶片和茎段在ＢＡ为０．５ｍｇ／Ｌ和ＮＡＡ为０．５ｍｇ／Ｌ的培养基上诱导产生愈伤组织,并在含０．２５ｍｇ／ＬＢＡ和０．５ｍｇ／ＬＮＡＡ的培养基上继代增殖。ＢＡ为０．５ｍｇ／Ｌ和ＮＡＡ为０．１ｍｇ／Ｌ可诱导叶片和愈伤组织发生不定芽,诱导频率分别为１００％和８２．９％,对于茎段,ＢＡ和ＮＡＡ分别为０．１ｍｇ／Ｌ和０．０１ｍｇ／Ｌ时诱导不定芽频率可达８３％。试管苗在大量元素减半并附加０．０１５ｍｇ／ＬＮＡＡ的ＭＳ培养基上诱导生根,生根率达８６．２％。     

Role of Exogenous Hormones in Inducing Cells in different Positions of Perianth Explants to Regenerate Flower Buds in Hyacinthus orientalis

Role of the exogenous hormone in inducing different position cells of perianth explants of hyacinth to regenerate flower buds was studied. Experiments showed that (1) Exogenous hormones are necessary for inducing cells of the perianth explant to regenerate the flower buds; (2) Only cytokinine alone could induce the regeneration of the flower buds, the auxin was not necessary; (3) Exogenous hormones in different concentrations could induce cells in the different parts of the perianth explants to differentiate the flower buds: 6-BAP or zeatin 2 mg/L alone could induce cells located at the lower part of the perianth to differentiate flower buds. Combination of 6-BAP or zeatin 2 mg/L and 2, 4-D 0.1 mg/L was advantageous to cells located middle part of the perianth to regenerate the flower buds. Combination of 6-BAP or zeatin 2 mg/L and 2, 4-D 1.0 mg/L could promote cells located at the upper part of the perianth to differentiate flower buds.     

中国木薯栽培种通过器官发生再生植株研究

本文研究了中国木薯栽培种四种外植体通过器官发生再生植株的条件。结果表明:在MS附加0.05mg/L TIBA,1mg/L BA的培养基上“NZ 188”初步的萌发胚状体“切头”后切口处可直接产生丛芽,出芽率为43%。“SC201”胚状体子叶块在MS附加0.5 mg/L NAA,0.5mg/L BA的培养基上可直接出芽,出芽率为42%,在MS附加0.5mg/L IBA,1.5mg/L BA培养基上·出芽率为31%,AgNO_3和ABA单独使用或配合使用均不利于芽的再生。“NZ188”胚状体下胚轴在MS附加0.5mg/LNAA,0.5mg/L BA的培养基上形成的愈伤组织转入MS附加1mg/L NAA,2mg/L BA的培养基上,3周后大多数愈伤组织有绿点出现、仅4.4%外植体分化出芽。“HZ188”无菌苗茎段接种在MS附加0.05mg/L TIBA,2mg/LBA的固体培养基上,2周后形成大量愈伤组织,4周后仅见一块愈伤组织分化出芽。