基于远缘杂交技术的海南野生兰花种质创新研究
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摘要
本研究以海南野生兰为材料,较为系统地了解其观赏特性、远缘杂交亲和性以及研究花粉贮藏及其生活力检测、人工授粉、远缘杂交种子无菌播种等远缘杂交育种技术,在此基础上进行种质创新,为创新利用海南丰富的热带兰花资源提供育种技术和材料。所获得的结果如下:
1对海南野生兰科植物观赏性状进行了较为全面的观察和评价。按观赏特性可分为大花型(24种)、多花型(23种)、长花序型(16种)、特长花序型(8种)、圆锥花序型(12种)、具香气型(20种)六类型。同时筛选了62种可直接开发利用的原生种,并从观赏角度将其划分为四类:观花类(15种)、观叶类(3种)、花叶兼具类(27种)、株型类(17种)。
2首次建立了较为完整的海南野生兰花远缘杂交技术体系,包括人工授粉技术、花粉块贮藏及其生活力检测技术、真假远缘杂种形态鉴别及其技术。花粉块经干燥a~ch后贮藏于Y℃下1年,采用花粉形态法、离体萌发法、授粉结实检测法检测,仍具有生活力。形态鉴别法根据子房膨大天数,柱头,杂交果大小、形态及颜色,种子颜色和形态,F1小苗性状等变化和差别,对真假杂种进行简单、直观地判断。
3建立了海南野生兰花远缘杂交种子无菌播种技术。获得以“花宝一号”、“MS”为基础,适当添加2.0~4.0 mg/L 6-BA、0.1~0.5 mg/L NAA及5~10%椰汁、香蕉泥等培养基,适于海南野生兰花种子无菌播种,已对33个种和30个杂交组合的种子进行了培养,并获得自交种苗及部分远缘杂种F1小苗。
4国内首次探讨了海南野生兰远缘杂交亲和性。对海南野生兰花进行大范围属、种间远缘杂交,涉及了海南现有兰科植物的34个属70个种,并获得部分远缘杂交种。种间杂交亲和性由高到低依次为毛兰属>隔距兰属>兰属>盆距兰属>石斛属>万代兰属。其中隔距兰属、毛兰属种间正、反交亲和性高。不同属间杂交亲和性由高到低依次是脆兰属>蜘蛛兰属>花蜘蛛兰属>虾脊兰属>尖囊兰属>带唇兰属>鹤顶兰属>五唇兰属、地宝兰属>毛兰属>美冠兰属>兰属>贝母兰属>万代兰属>石豆兰属>金石斛属>血叶兰属>隔距兰属>石斛属>钻喙兰属等。
5获得创新种质12份。筛选了20个具有优良性状的亲本组合,11个组合已萌发,7个获得一定数量远缘杂种F1小苗,并移栽成活,长势良好。国内首次通过远缘杂交获得叶缘金黄的变异类型。
In this study, some wild orchids came from Hainan Province were used for materials. For the purpose of systemically learning the distant-hybridization technique, e.g. cornamental haracteristics, distant hybridization’s cross compatibility, pollinium storage and testing of viability, artificial pollination, asepsis sowing and so on were studied. Besides, for utilizing abundant resources and offering technology and materials of Hainan Province’s orchids, germ plasm enhancement were progressed in the base of early period work.The main results and conclusion are aummarized as follows:
1 Systemically observation and assessment on the cornamental haracteristics of wild orchids in Hainan Province. They were classified in 6 types according to enjoyment characteristic, viz. big flowers(24 species), many flowers (23 species), long inflorescence(16 species) , even longer inflorescence(18 species) , panicle(12 species) and fragrance (20 species). Simultaneity , 62 initial species were screening for development and utilization direct. From the gardens enjoyment angle, they can be divided into 4 types: flowering type(15 species), foliage plant (3 species),flowering & foliage plant (27 species) and plant type(17 species).
2 The first completely full technology system of distant hybridization for wild orchids that in Hainan had been built up.It included techniques as artificial pollination, pollinium storage and their viability testing, morphological identification of true or not hybrids. The pollinium storage and viability testing technique: the pollinium were storaged under Y℃after drying a~ch and the pollen were still alive after 1 year which were tested by morphological identification, germination in vitro and pollination. The morphological identification method:through the change of column, days of ovary’s swell, the size,the shape and the color of the hybrid fruit,the shape and the color of the seeds,the change and distinguish of the young F1 characteristics to judge the offspring was a hybrid or not simply and quickly.
3 The asepsis sowing technique for the hybrids of distant hybridization of the wild orchids in Hainan was astablished. The mediums of“Hy-NO.1”and“MS”containing 2.0~4.0 mg/L 6-BA ,0.1~0.5 mg/L NAA and supplemented with 5~10% coconut milk or banana mud were fit for asepsis sowing of the wild orchids in Hainan.33 species and 30 hybrids of distant-hybridization had been brought up well in the two mediums. Some selfed seedling and young F1 plantlets had been grew well.
4 The first study on the cross compatibility of Hainan’s wild orchids in China was discussed. The study on interspecific hybridization and intergneric hybridization in large scale of wild orchids in Hainan were involved with 34 genus and 70 species of wild orchids in Hainan existing and some hybrids were obtained. The cross compatibility of interspecific hybridization from strong to weak were Eria, Cleisostoma, Cymbidium, Gastrochilus, Dendrobium, Vanda. The cross compatibility of reciprocal cross in Cleisostoma and Eria were stronger. The cross compatibility of intergneric hybridization from strong to weak were Acampe, Arachnis, Calanthe, Kingidium, Tainia, Phaius, Doritis, Geodorum, Eria, Eulophia, Cymbidium, coelogyme, Vanda, Bulbophyllm, Cleisostoma, Dendrobium, Rhymchostylis and so on.
5 Twelve germplasm enhancements had been obtained. Twenty combinations with eximious characteristics were screened for germplasm enhancement. Eleven combinations had germianted and seven combinations had some young F1 with well transplantating and surviving.The first variant type with yellow leaf margin by distant hybridization had been obtainted.
1对海南野生兰科植物观赏性状进行了较为全面的观察和评价。按观赏特性可分为大花型(24种)、多花型(23种)、长花序型(16种)、特长花序型(8种)、圆锥花序型(12种)、具香气型(20种)六类型。同时筛选了62种可直接开发利用的原生种,并从观赏角度将其划分为四类:观花类(15种)、观叶类(3种)、花叶兼具类(27种)、株型类(17种)。
2首次建立了较为完整的海南野生兰花远缘杂交技术体系,包括人工授粉技术、花粉块贮藏及其生活力检测技术、真假远缘杂种形态鉴别及其技术。花粉块经干燥a~ch后贮藏于Y℃下1年,采用花粉形态法、离体萌发法、授粉结实检测法检测,仍具有生活力。形态鉴别法根据子房膨大天数,柱头,杂交果大小、形态及颜色,种子颜色和形态,F1小苗性状等变化和差别,对真假杂种进行简单、直观地判断。
3建立了海南野生兰花远缘杂交种子无菌播种技术。获得以“花宝一号”、“MS”为基础,适当添加2.0~4.0 mg/L 6-BA、0.1~0.5 mg/L NAA及5~10%椰汁、香蕉泥等培养基,适于海南野生兰花种子无菌播种,已对33个种和30个杂交组合的种子进行了培养,并获得自交种苗及部分远缘杂种F1小苗。
4国内首次探讨了海南野生兰远缘杂交亲和性。对海南野生兰花进行大范围属、种间远缘杂交,涉及了海南现有兰科植物的34个属70个种,并获得部分远缘杂交种。种间杂交亲和性由高到低依次为毛兰属>隔距兰属>兰属>盆距兰属>石斛属>万代兰属。其中隔距兰属、毛兰属种间正、反交亲和性高。不同属间杂交亲和性由高到低依次是脆兰属>蜘蛛兰属>花蜘蛛兰属>虾脊兰属>尖囊兰属>带唇兰属>鹤顶兰属>五唇兰属、地宝兰属>毛兰属>美冠兰属>兰属>贝母兰属>万代兰属>石豆兰属>金石斛属>血叶兰属>隔距兰属>石斛属>钻喙兰属等。
5获得创新种质12份。筛选了20个具有优良性状的亲本组合,11个组合已萌发,7个获得一定数量远缘杂种F1小苗,并移栽成活,长势良好。国内首次通过远缘杂交获得叶缘金黄的变异类型。
In this study, some wild orchids came from Hainan Province were used for materials. For the purpose of systemically learning the distant-hybridization technique, e.g. cornamental haracteristics, distant hybridization’s cross compatibility, pollinium storage and testing of viability, artificial pollination, asepsis sowing and so on were studied. Besides, for utilizing abundant resources and offering technology and materials of Hainan Province’s orchids, germ plasm enhancement were progressed in the base of early period work.The main results and conclusion are aummarized as follows:
1 Systemically observation and assessment on the cornamental haracteristics of wild orchids in Hainan Province. They were classified in 6 types according to enjoyment characteristic, viz. big flowers(24 species), many flowers (23 species), long inflorescence(16 species) , even longer inflorescence(18 species) , panicle(12 species) and fragrance (20 species). Simultaneity , 62 initial species were screening for development and utilization direct. From the gardens enjoyment angle, they can be divided into 4 types: flowering type(15 species), foliage plant (3 species),flowering & foliage plant (27 species) and plant type(17 species).
2 The first completely full technology system of distant hybridization for wild orchids that in Hainan had been built up.It included techniques as artificial pollination, pollinium storage and their viability testing, morphological identification of true or not hybrids. The pollinium storage and viability testing technique: the pollinium were storaged under Y℃after drying a~ch and the pollen were still alive after 1 year which were tested by morphological identification, germination in vitro and pollination. The morphological identification method:through the change of column, days of ovary’s swell, the size,the shape and the color of the hybrid fruit,the shape and the color of the seeds,the change and distinguish of the young F1 characteristics to judge the offspring was a hybrid or not simply and quickly.
3 The asepsis sowing technique for the hybrids of distant hybridization of the wild orchids in Hainan was astablished. The mediums of“Hy-NO.1”and“MS”containing 2.0~4.0 mg/L 6-BA ,0.1~0.5 mg/L NAA and supplemented with 5~10% coconut milk or banana mud were fit for asepsis sowing of the wild orchids in Hainan.33 species and 30 hybrids of distant-hybridization had been brought up well in the two mediums. Some selfed seedling and young F1 plantlets had been grew well.
4 The first study on the cross compatibility of Hainan’s wild orchids in China was discussed. The study on interspecific hybridization and intergneric hybridization in large scale of wild orchids in Hainan were involved with 34 genus and 70 species of wild orchids in Hainan existing and some hybrids were obtained. The cross compatibility of interspecific hybridization from strong to weak were Eria, Cleisostoma, Cymbidium, Gastrochilus, Dendrobium, Vanda. The cross compatibility of reciprocal cross in Cleisostoma and Eria were stronger. The cross compatibility of intergneric hybridization from strong to weak were Acampe, Arachnis, Calanthe, Kingidium, Tainia, Phaius, Doritis, Geodorum, Eria, Eulophia, Cymbidium, coelogyme, Vanda, Bulbophyllm, Cleisostoma, Dendrobium, Rhymchostylis and so on.
5 Twelve germplasm enhancements had been obtained. Twenty combinations with eximious characteristics were screened for germplasm enhancement. Eleven combinations had germianted and seven combinations had some young F1 with well transplantating and surviving.The first variant type with yellow leaf margin by distant hybridization had been obtainted.
引文
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