Studies on induction of multiple buds and adventitious embryo in Shorea spp.of Dipterocarpaceae

• Project/Area Number: 16580129
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: 林産科学・木質工学
• Research Institution: Tokyo University of Agriculture and Technology
• Principal Investigator: KUBO Takafumi Tokyo University of Agriculture and Technology, Institute of Symbiotic Science and Technology, Professor, 大学院・共生科学技術研究部, 教授 (00015091)
• Co-Investigator(Kenkyū-buntansha): OGITA Shinjiro Toyama Prefecture University, Faculty of Technology, Lecturer, 工学部, 講師 (50363875) ; 中村 健太郎 住友林業(株), 筑波研究所, 研究員
• Project Period (FY): 2004 – 2005
• Project Status: Completed (Fiscal Year 2005)
• Budget Amount: ¥3,600,000 (Direct Cost: ¥3,600,000); Fiscal Year 2005: ¥1,600,000 (Direct Cost: ¥1,600,000); Fiscal Year 2004: ¥2,000,000 (Direct Cost: ¥2,000,000)
• Keywords: Multiple bud / Adventitious embryo / Diperocarpaceae / Shorea multiflora / Callus / Plant regulators / CO_2 exposure / MS medium / フタバガキ科樹種 / 多芽体誘導 / カルス誘導 / 細胞形態 / シュート化 / 発根 / 初期不定胚 / 植物調整物質

Research Abstract

1. Examination of culture conditions for inducing multiple buds from apical shout and lateral bud
It was attempted to induce the multiple buds from apical shout of Shorea.multiflora by rotation culture in liquid medium, by relating to effects of various culture conditions, as MS medium and plant regulators, which were auxin of 2,4-D and cytokinin of BAP, zeatin and 4PU. Scale leaves were formed by exclusive application of BAP 3μM, but not by other conditions. The elongation was recognized by 25% of frequency, but these shouts died within a month. In this experiment, although multiple buds were not induced, it was suggested that cytokinin was effective for the formation of multiple buds.
2. Examination of culture conditions for callus induction, evaluation of the differentiation potency and culture conditions for formation of early adventitious embryo
Induction of callus with the differentiation potency was tried using leaves of S.multiflora, by relating to the effects of plant regulators of auxin (2,4-D) and cytokinin (BAP, 4PU, zeatin and TDZ), two kind of sugars (sucrose and maltose) and exposure of CO_2. The combination of 10μM of 2,4D and 3μM of BAP was the most effective for inducing frequently the excellent callus. Furthermore, the developed tissues like apical meristem or primary globular embryo was observed in the induced callus. Also, it was recognized that maltose, TDZ and exposure of CO_2 were useful for callus proliferation.
3. Examination of culture conditions for shouting and rooting of the induced multiple buds
In this assignment, an effective result was not achieved for the study plan. As for rooting, it may need to examine in detail regarding to the modification of basic medium and the improvement of the concentration and combination of plant regulators in the future.
4. Examination of culture conditions for maturation of primary embryo
It was attempted to maturate the tissue like a primary globular embryo, observed in the induction callus. Actually, the effects of Auxin and cytokinin were investigated to promote the maturation, but the development was not recognized in every culture conditions.