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標題: 紫錐菊(Echinacea purpurea (L.) Moench)花藥培養
Anther culture of purple coneflower (Echinacea purpurea (L.) Moench)
作者: 彭筱娟
Peng, Hsiao-Chuan
Contributors: 陳宗禮
農藝學系所
關鍵字: 紫錐菊;花藥培養
purple coneflower;anther culture
日期: 2013
Issue Date: 2013-11-21 11:05:12 (UTC+8)
Publisher: 農藝學系所
摘要: 紫錐菊 (Echinacea purpurea) 為新興保健及觀賞作物,極具市場生產價值,但紫錐菊為異交作物,且具自交不和合性,若欲快速育成新品種,利用組織培養技術培育同質二倍體,做為育種材料是理想的方法。本研究以花藥進行癒傷組織誘導,選用具單核期小孢子之紫錐菊花藥,接種於以1/2 MS為基礎培養基,配合施用18種不同NAA與BA濃度組合之花藥誘導癒傷組織培養基,經8週培養後,發現各組合皆可成功誘導癒傷組織,惟不同處理組合間的癒傷組織具明顯差異,其中當NAA 1.00 mg L-1 及 BA 0.40 mg L-1之組合具最佳花藥癒傷組織誘導率,且在不同BA濃度處理間,具顯著性差異;當NAA 1.00 mg L-1 及 BA 0.40 mg L-1之組合具最佳花藥癒傷組織增殖率,且在不同NAA濃度處理間,具顯著性差異。花藥培養期間,以花藥囊開裂為標準,進行早期篩選;後期挑選生長良好、質地鬆散癒傷組織,進一步以不同階段的液態培養方式誘導芽體產生。在配合蔗糖、葡萄糖的施用與不同NAA、BA比例組合處理下,以癒傷組織直接誘導芽體,各處理之芽體誘導效果皆不理想;若花藥癒傷組織先以液態培養基進行增殖後再誘導芽體產生,雖延長培養時間但較能有效獲得芽體。為判別花藥培養誘導株是否自花藥小孢子發育,隨機挑選75株誘導株,取花藥供給親做為對照,進行ISSR分子檢定技術分析親源後,再以根尖染色體壓片確認誘導株倍數性,其中6株為雙倍體,2株為鑲嵌體 (單+雙倍體)。
The pure line of homozygous double haploid plant provides the other possibility in plant breeding, especially for the hybrid plant. In this study, we used purple coneflower (Echinacea purpurea) anthers which containing microspores at uninucleate stage as culture material. Anther were cultured on basal medium (1/2 MS medium) consisting various NAA and BA concentrations. During the 18 different dose of NAA and BA, callus formation percentage were significant differences among various BA concentrations, the highest callus formed rate reached 96.6%, when anther were cultured on the medium supplemented with NAA 1.00 mg L-1 and BA 0.40 mg L-1. Callus length also were significant differences among various NAA concentrations, the longest callus length was 5.07 mm , when anther were cultured on the medium contain NAA 1.00 mg L-1 and BA 0.80 mg L-1. The regenerated plantlets were generated from anther callus when they were pre-culture by suspension. The embryogenic cell from anther culture were incubated in different way, proliferated callus before induce regenerated plantlets increase the culture period, but it is more efficiently to induce large number of cotyledon-like embryo. For selecting the plantlets from the microspores, ISSR analysis has been tried. 75 regenerated plantlets were randomly selected from anther culture for ISSR analysis, and amplified polymorphic bands between anther donor plant and anther culture induced plantlets. In chromosome number examination of root-tip cell of regenerated plantlets, there are 6 of them are diploid and 2 of them are chimera (haploid and diploid).
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