拖鞋蘭微體繁殖最適條件探討

Optimization of Paphiopedilum micranthum ‘Viet’ regeneration for industrial application

武晉平(Binh-Vu Tien)

指導教授 ： 戴守谷

國立虎尾科技大學/文理學院/生物科技系碩士班/碩士(2017年)

摘要

Paph. micranthum 屬於拖鞋蘭亞科，越南北部到廣西西北部。研究目的是確定不同濃度的 2,4-D（2,4-二氯苯氧基乙酸）組合與 BAP（6-芐基氨基嘌呤）和蔗糖濃度對來自葉部外植體的芽體和根再生的影響，以成功繁殖拖鞋蘭 ，並用於產業發展。
拖鞋蘭切成 (0.5 x 0.5 cm) 外植體培養在 15 毫升的 MS 培養基沒有植物生長調節劑作為對照組與不同含量 2,4-D 和 BAP，包含: 0.5、1.0、2.0 ppm。根 2-3 mm和葉誘導癒傷組織在不含 PGRs 和 2,4-D 組合 BAP 的 1/2 MS 培養基。結果表明：在接種後 4 週內，2.0ppm 2,4-D 和 0.5ppm BAP 組合的 1/2 MS 培養基上培養時，從外植體獲得癒傷組織誘導率高達 93.3％。此外，從具有 2.0ppm 2,4-D及 2.0ppm BAP 的 1/2 MS 培養基上葉的癒傷組織有最佳的芽體再生。大約 86.6％的誘導培養為 3.3 個芽體及 3.0 個葉在每一個外植體。2,4-D 組合 BAP 的濃度在培養 4 個月後每個外植體誘導根的數量和長度為2.1 根/外植體和長度為 1.1 (cm)。在加入有 30g/L蔗糖的 1/2 MS 培養基上葉數和根數最多，其中 3.8 葉/外植體，3.3 根/外植體，根長 1.8（cm）。在 40g/L 蔗糖下，幼苗的發育減少。最低的是沒有蔗糖的培養基。這項研究表明，2.0 ppm 2,4-D與 2.0 ppm BAP 的組合有效地誘導了拖鞋蘭 (Paph. micranthum) 芽和根。蔗糖濃度對 Paph. Micranthum 再生有重要的影響。濃度為 30g/L 蔗糖的 1/2 MS 培養基上，對幼苗的發育生長是最佳的。

關鍵字

Paph. Micranthum ； 再生 ； 植物生長調節劑(PGR) ； 1/2 MS 培養基 ； 蔗糖濃度

並列摘要

Paphiopedilum micranthum ‘Viet’ (Paph. micranthum‘Viet’), also called‘Viet’, belongs to the subfamily Cypripedioideae of the family Orchidaceae, is a new strain of Paph. micranthum from northern Vietnam to south of the western in China. The present study examined the effects of various concentrations of 2,4-Dichlorophenoxyacetic acid (2,4-D) combined with 6-benzylaminopurine (BAP) and sucrose concentrations on the regeneration of shoot and root from leaf explants of Paph. micranthum ‘Viet’ for industrial application.
Young fresh leaves of Paph. micranthum ‘Viet’ were cut into small explants (0.5 x 0.5 cm) and were cultured in glass tubes containing 15 mL of a medium developed by Murashige and Skoog (MS medium) at half strength both
without plant growth regulators (control medium) and containing combinations of 2,4-D and BAP of varying concentrations (0.5, 1.0, and 2.0 ppm). Shoot tip 2-
3 mm from callus induction and rootless shoot with 2 leaves were transferred vertically on half-strength MS medium without plant growth regulators and 2,4-
D combined BAP to optimize shoot and root induction. Rootless shoot with 2 leaves were transferred vertically on half-strength MS medium with different sucrcose concentrations (0, 10, 20, 30, 40 g/L) to optimize on the growth and development of seedling.
The results showed that a higher proportion of callus induction (93.3%) was observed in leaf explants when cultured on half-strength MS medium supplemented with 2.0 ppm 2,4-D + 0.5 ppm BAP combination for 4 weeks of
inoculation. Furthermore, the optimum degree of shoot regeneration from the leaf-derived callus was placed on half-strength MS medium with 2.0 ppm 2,4-D
+ 2.0 ppm BAP. Approximately 86.6% of cultures responded with an average number of 3.3 shoots and 3.0 leaves/explant. In addition, the combination of 2,4-
D and BAP of equal concentrations provided the optimal number and length of
roots per explant after 4 months of culture, which were 2.1 roots per explant and
1.1 cm in length, respectively. On half-strength MS medium supplemented with
30 g/L sucrose was the highest number of leaves and roots and also length of
root, which gave 3.8 leaves/explant, 3.3 roots/explant, 1.8 (cm) length of root.
At 40 g/L sucrose gave to decrease in the development of seedling. The lowest
was the medium without sucrose.
The findings of the present study indicate that BAP combined with 2,4-D
effectively induces the growth of shoots and roots in Paph. micranthum ‘Viet’.
Sucrose positively affected on the characteristics of Paph. micranthum ‘Viet’.
We could be estimated that on half-strength MS medium supplemented with 30
g/L sucrose was the optimal on the growth of seedling developed.

並列關鍵字

Paphiopedilum micranthum ； Regeneration ； Plant growth regulators (PGRs) ； Half-strength MS medium ； Sucrose concentration

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