培養基鹽類配方、活性碳與蔬果添加物對台灣白及無菌播種之影響

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(1)ѯᢋҪІณຄክᆎ!. ѯᢋၼཿः‫( ف‬J. Taiwan Agric. Res.) 60(2):77–88 (2011). 77. ୻Ꭶ୷ᡶᜪଛБǵࢲ‫܄‬ᅹᆶጫ݀బу‫ނ‬ჹѠ᡼қϷค๵ 1 ኞᅿϐቹៜ ఆ຾က2 ഋ࠶Խ2 হ‫✔ڻ‬2,3! ᄣ. ्. కໍဏȃചࡅՌȃৈ‫ۊ‬⢐Ȅ2010ȄஊᎵஆᢅ᜹ପРȃࣁ‫ܓ‬ᆈᇅጺ‫఻ݏ‬ђ‫ޑ‬ᄈѯᢋҪІณຄ ክᆎϟኈ៫Ȅѯᢋၼཿः‫ ف‬60:77–88Ȅ. Ґः‫ف‬ᙥҦ 5 ᆎϛӤᢅ᜹ପР Kundson (Knudson 1946)ȃVW (Vacin & Went 1949)ȃӓ໕Іљ໕ϟ MS (Murashige & Skoog 1962) пІ߇᝙ 1 ဵ (HYPONeX 7-6-19) ࣲ఻ђ 0.8% Bacto agar ᇅ 3%ጶᑦ࣐ஆᙄஊᎵஆȂପӬ 0.1%ࣁ‫ܓ‬ᆈȂпІ ϛӤ֥໕ϟॸᒃ‫଼ܗ‬ႜᖧ‫఻ޠ‬ђȂӔ 4 ᆎၑᡜໍ՘ѯᢋҪІ༂ዤ‫ݏ‬ುณຄክᆎȂ ขၑαख़ӱφᄈѯᢋҪІᆎφึ߄ᇅߵໍ߄ᡞҢߞึ‫ي‬ϟኈ៫Ȅၑᡜ๗‫ݏ‬ᡘұ‫ڦ‬ Ս༂ዤ‫ݏ‬ುϟᆎφӶณຄክᆎȂஊᎵܼ (25 ± 1)кᕘძ 8 Љࡤ֊Ѡึ߄ȂஊᎵ 50 ЉࡤᡘұяӶ 5 ᆎϛӤᢅ᜹ପРᇅ֥ԥ 0.1%ࣁ‫ܓ‬ᆈȃॸᒃ‫଼ܗ‬ႜᖧጺ‫఻ݏ‬ђ‫ޑ‬ (2.5ȃ5.0ȃ7.5ȃ10%) ϟஊᎵஆϜȂᄈᆎφึ߄ኈ៫ϛᡘ຀ (P > 0.05)Ȃϊ़൑ੂ ᘁ२ܼ 1/2 MS ఻ђ 0.1%ࣁ‫ܓ‬ᆈஊᎵஆϜѠႁ 20.0 mgȂܼ MS ఻ђ 0.1%ࣁ‫ܓ‬ᆈஊ Ꮅஆ࣐ 20.8 mgȂՅܼ VW ఻ђ 0.1%ࣁ‫ܓ‬ᆈஊᎵஆ༊ 7.4 mgȂп 1/2 MS ପРஊᎵ ஆᄈϊ़ึ‫ي‬ϟߵໍਞ‫ݏ‬ഷᡘ຀ȇ఻ђࣁ‫ܓ‬ᆈԥֆܼϊ़ึ‫ي‬ȇ఻ђॸᒃ‫଼ܗ‬ႜ ᖧࣲѠߵໍϊ़ᘁ२‫ޠ‬ቩђȂՅп଼ႜᖧၷॸᒃ࣐‫ٺ‬Ȃ‫఻ڐ‬ђϟഷᎍᐩ࡚࣐ 7.5%Ȅ ӱԫȂп 1/2 MS ᢅ᜹ȂପӬ 0.1%ࣁ‫ܓ‬ᆈᇅ 7.5%଼ႜᖧ‫఻ޠ‬ђ࣐ഷ‫ٺ‬ѯᢋҪІณ ຄክᆎᇅஊᎵϟஊᎵஆȄ ᜱᗥມȋѯᢋҪІȃᆎφึ߄ȃϊ़ึ‫ي‬ȃ౮ϲஊᎵȄ. ࠊ. ّ. ѯ ᢋ Ҫ І [Bletilla formosana (Hayata) Schltr.] ࣐ѯᢋ੬ԥᆎϟ៍ऌෛ‫ޑ‬Ȃϸҁܼѯᢋ ӓძϟ҂ӵᇅାύӵୣȂᇅϜ᛿؆‫ܛ‬ңϟҪІ [B. striata (Thunb) Reichb. f.] ࣐Ӥ឵౵ᆎȂ‫ڐ‬ ༶ೂҼѠձ࣐᛿؆‫ٻ‬ңȂෛੂ‫ٯ‬চ‫ڏ‬༫᛻ᢏ፭. ቌঅ (Na et al. 1978)ȄҪІл्пϸੂ‫ݳ‬ᖆ ෦Ȃᖆ෦ഁ౦ጦᄛȂณ‫ݳ‬ᅗ٘೤ዂϾᆎෛ‫ޠ‬ሰ ؒ (Lin et al. 1994; Yang et al. 2002)Ȃйпณ‫ܓ‬ ϸੂРԓᖆ෦Ȃᆎ़ܿ‫ࢴ੿ڨ‬ད࢘Ȃ‫ٯ‬ԥᆎ‫ܓ‬ ଞϾϟુᘉ (Yang et al. 2002; Zhang et al. 2005)ȇԫѵӱ࣐ҪІ঩Ңӵ‫ޠ‬෶Ўᇅ᎐‫ڨ‬ખ ᚾȂ‫ٻ‬ூѯᢋҪІᇅϜ୾τഛҪІϟഐҢఋထ. 1. ՘࢈ଲၼཿ‫ې‬সཽၼཿၑᡜ‫ܛ‬ः‫ف‬ൣ֚ಒ 2502 ဵȄ௦‫ڨ‬С෉Ȉ100 Ԓ 3 У 31 СȄ 2. Ґ‫ܛ‬Ң‫׭ޑ‬೛ಣဓңֆ౪ः‫ف‬সȃֆ౪ः‫ف‬সІ୚ः‫ف‬সȄѯᢋ ѯϜҀȄ 3. ೾ଊձ޲ȂႬφ໐ӈȈ[email protected]ȇ༉઎ᐡȈ(04)23302806Ȅ.

(2) ѯᢋၼཿः‫ف‬. 78. ҔӶ‫ץ‬ഁ෶ЎϜ (Lin et al. 1994; Zhu & Wang 1999; Fu et al. 2006)ȂϜ୾ϑ஡ҪІӗ࣐ࣣไᛕ ӡ‫ߴޠ‬ៗෛ‫( ޑ‬Peng et al. 2004)Ȃ‫ٯ‬໡ۗпಣ ᙒஊᎵРԓໍ՘ᖆ෦ः‫ف‬ȂԚਞࣻ࿌ًԂ (Peng et al. 2004; Yang et al. 2002; Zhang et al. 2005)ȄѯᢋҪІӶณຄክᆎР८ԟ෉ԥЎ໕ϟ ः‫ف‬ȂLin et al. (1994) ࡿяѯᢋҪІпथึ‫ي‬ ෉ՎനႫࠊ෉ (torpedo) ϟᆎφ຃߄౦ഷ‫ٺ‬Ȃ ‫ٯ‬ѠւңᚗੋЬໍ՘ᆎφႲ௪೏౪Ȃႁ‫ژ‬ණԟ ึ߄ϟਞ‫ݏ‬ȄYeh et al. (1995) ‫ޠ‬ः‫ࡿࠍف‬яȂ MS (Musashiage & Skoog 1962) ஆҐᢅ᜹Іጶ ᑦ࣐ණ‫ٽ‬ѯᢋҪІ঩౩ᡞึ‫्҇ي‬ϟᕋᎵԚ ϸȂйစҦ peptoneȃNAA ‫఻ޠ‬ђпІପӬӏ ྲ೏౪Ѡߵໍϊ़ϟึ‫ي‬ȄѯᢋҪІ࣐ѯᢋҐ ρ঩Ңෛ‫ޑ‬ȂѠձ࣐᛿؆І༫᛻ᢏ፭ϟңȂՄ ໕‫ڐ‬ഐҢఋထҔഃᅛ෶ЎȂঅூ֏΢Ӷᆎ঩ߴ ԇІᆎ़ᖆ෦Р८ђ஽ः‫ف‬໡ึȄҐः‫ف‬пϛ Ӥᢅ᜹ପРஊᎵஆପӬࣁ‫ܓ‬ᆈȃॸᒃᇅ଼ႜᖧ ๊ጺ‫ݏ‬ԥᐡ‫఻ޠޑ‬ђȂᑣᒶяᄈܼѯᢋҪІณ ຄክᆎІ౮़Ңߞഷ‫ٺ‬ϟஊᎵஆಣԚȂ࡛Ҵѯ ᢋҪІಣᙒஊᎵτ໕ᖆ෦ᡞ‫ق‬Ȅ. ؆ਠᇅР‫ݳ‬ ၑᡜ؆ਠᇅ੒ࢴ Ґः‫ٻܛف‬ңϟѯᢋҪІ௵Սۤ៍ង߇ϵ ၰȂҦ߇ጹ‫ً׾‬ൠ៍໩ϸൠ‫ܛ‬ණ‫ٽ‬Ȅෛੂ໡߇ ࡤп΢ϏՍ߇௳લȂစ 120 Љࡤ‫ۧڦ‬ґ໡ບϟ ༂ዤ‫ݏ‬ುձ࣐ณຄክᆎϟңȄ‫ݏ‬ುӒп 70%଩ ᆡໍ՘ߓ८੒ࢴȂӕܺΤ఻ђ Tween 20 ৥຀Ꮩ ϟ 0.6%Ԫ෪ሗ໙ྚషϜȂпົॲ‫ݱ‬Ꭻᕞ 1 ϸ យȂӕпК཯Ꭻᕞ੒ࢴ 15 ϸយȂ‫ࡤڐ‬пณຄЬ ఽࣀ 3 ԪȂ੒ࢴࡤϟ‫ݏ‬ುп၍ঢΧঢ໡ࡤ‫ڦ‬я ᆎφໍ՘ክᆎȂ‫ໍٯ‬՘ϛӤஆҐᢅ᜹ȃ఻ђ 0.1%ࣁ‫ܓ‬ᆈϟϛӤஆҐᢅ᜹ȃॸᒃԥᐡ఻ђ‫ޑ‬ ପӬ఻ђ‫ܗ‬ϛ఻ђ 0.1%ࣁ‫ܓ‬ᆈȃ଼ႜᖧԥᐡ఻ ђ‫ޑ‬ପӬ఻ђ‫ܗ‬ϛ఻ђ 0.1%ࣁ‫ܓ‬ᆈᄈѯᢋҪ Іᆎφึ߄ᇅϊ़ึ‫ي‬ϟኈ៫๊Ӕ 4 ᆎၑᡜȄ. ಒ 60 ‫ڣ‬. ಒ2෉. ஆҐᢅ᜹ପРஊᎵஆϟኈ៫ၑᡜ೪ॏ ณຄክᆎ‫ٻܛ‬ңϟஊᎵஆ‫ڐ‬ஆҐᢅ᜹ପР ϸր࣐ Kundson (Knudson 1946)ȃVW (Vacin & Went 1949)ȃӓ໕Іљ໕ MS (Murashige & Skoog 1962) пІ߇᝙ 1 ဵ (HYPONeX 7-6-19, USA) Ӕ 5 ᆎȂ࣐൑ӱφၑᡜȂө೏౪ 5 २ፓȂ п௥ଇϛӤஆҐᢅ᜹ᄈѯᢋҪІᆎφึ߄ᇅϊ ़ึ‫ي‬ϟኈ៫Ȃ‫ڐ‬Ϝ VW І MS ࣐લҒ୧ࠣ (Duchefa, Netherlands)Ȅ‫ܛ‬ԥପРஊᎵஆࣲ఻ђ 3% (30 g·L-1) ጶᑦІ 0.8% (8 g·L-1) Bacto agar (Difco, USA)Ȅ. 0.1%ࣁ‫ܓ‬ᆈ఻ђϟኈ៫ၑᡜ೪ॏ пαख़ஊᎵஆ఻ђ൑Κ 0.1% (g·L-1) ࣁ‫ܓ‬ ᆈȂ௵ؑΚ೏౪ө 5 २ፓ‫ޠ‬൑ӱφၑᡜȂп௥ ଇࣁ‫ܓ‬ᆈ‫఻ޠ‬ђᄈѯᢋҪІᆎφึ߄ᇅϊ़ึ ‫ي‬ϟኈ៫Ȅ. ϛӤᐩ࡚ϟॸᒃԥᐡ఻ђ‫ޑ‬ପӬ఻ђ‫ܗ‬ϛ ఻ђ 0.1%ࣁ‫ܓ‬ᆈϟኈ៫ၑᡜ೪ॏ ၏Ρӱφၑᡜ࢑пαख़ 1/2 MS ᢅ᜹ପРஊ Ꮅஆϸր఻ђ 2.5% (25 g·L-1)ȃ5.0% (50 g·L-1)ȃ 7.5% (75 g·L-1)ȃ10% (100 g·L-1) ϟॸᒃȂ‫ٯ‬ପ Ӭ఻ђ‫ܗ‬ϛ఻ђ 0.1%ࣁ‫ܓ‬ᆈȂؑΚ೏౪ಣӬө 5 २ፓȂп௥ଇஊᎵஆϜϛӤᐩ࡚ϟॸᒃ఻ђ ‫ޑ‬ᇅ 0.1%ࣁ‫ܓ‬ᆈᄈѯᢋҪІᆎφึ߄ᇅϊ़ ึ‫ي‬ϟኈ៫Ȅ. ϛӤᐩ࡚ϟ଼ႜᖧԥᐡ఻ђ‫ޑ‬ପӬ఻ђ‫ܗ‬ ϛ఻ђ 0.1%ࣁ‫ܓ‬ᆈϟኈ៫ၑᡜ೪ॏ ၏Ρӱφၑᡜ࢑пαख़ 1/2 MS ᢅ᜹ପР ஊᎵஆϸր఻ђ 2.5%ȃ5.0%ȃ7.5%ȃ10%ϟ଼ ႜᖧȂ‫ٯ‬ପӬ఻ђ‫ܗ‬ϛ఻ђ 0.1%ࣁ‫ܓ‬ᆈȂؑΚ ೏౪ಣӬө 5 २ፓȂп௥ଇஊᎵஆϜϛӤᐩ࡚ ϟ଼ႜᖧ఻ђ‫ޑ‬ᇅ 0.1%ࣁ‫ܓ‬ᆈᄈѯᢋҪІᆎ φึ߄ᇅϊ़ึ‫ي‬ϟኈ៫Ȅ. ѯᢋҪІᆎφ௦ᆎᇅஊᎵᕘძʳ αख़ϟஊᎵஆପᇨ‫׈‬Ԛࡤ஡ϟ pH ࣲ።ᐍ Վ 5.6Ȃӕп 121ʚྟຄ 20 ϸយȄஊᎵϟၑᆔ.

(3) ѯᢋҪІณຄክᆎ!. (25 mm × 150 mm) ؑᆔဋ 10 mL ஊᎵஆȂӶ੒ ࢴࡤᘜԚఈ८տࠔറңȄณຄክᆎп௦ᆎᕘ‫ݯ‬ ‫ڦ‬ᆎφࡤȂ஡ᆎφ҂ְයဋܼஊᎵஆߓ८Ȃؑ Мၑᆔϲτङ௦ᆎ 200–250 ঐᆎφȄ௦ᆎࡤဋ ܼ (25 ± 1)кࡰྤȃӏྲ஽࡚ 38 Pmol·m-2·s-1 І 14 ϊਣӏ໋෉ᕘძϜஊᎵȄ. ಜॏϸ‫ݚ‬ αख़ 4 ᆎၑᡜ‫ܛ‬ூၦਠְစ SAS 9.1.3 ಜ ॏϸ‫ݚ‬৉၇೻ᡞ (SAS Institute Inc. 2002) ໍ՘ ᡑРϸ‫( ݚ‬ANOVA) ࡤȂस೏౪໣ৰ౵ᡘ຀Ȃ ࠍӕւң least significant difference (LSD) ขᡜ Щၷө೏౪҂ְঅ໣ϟঐրৰ౵‫ܓ‬Ȅ. ѯᢋҪІϊ़ึ‫ي‬ਣ෉ୣϸᇅ‫ޒܓ‬።ࢦʳ ࣐ຠզαख़өၑᡜ‫ޠ‬ϛӤஊᎵஆ೏౪ᄈᆎ φึ߄ᇅϊ़ึ‫ي‬ϟኈ៫Ȃ።ࢦਣ஡ϊ़ึ‫ي‬ ਣ෉ୣϸԚ 1 (঩౩ᡞ)ȃ2 (ಒΚбဩߞя)ȃ3 (ಒ ΚбဩԚዤߞяಒΡбဩ)ȃ4 (ಒΡбဩߞτܼ 0.5 cm)ȃ5 (ಒήбဩߞτܼ 0.5 cm) Ӕ 5 ঐਣ ෉ȂԄშ 1 ‫ܛ‬ұȂܼክᆎࡤ 50 Љໍ՘ึ߄౦ȃ ϊ़ึ‫ي‬ਣ෉ᇅϊ़ᘁ२๊‫ޒܓ‬።ࢦȄึ߄౦ ࣐ؑМၑᆔϜ঩౩ᡞ (֥) пαึ‫ي‬ਣ෉ϟᖃ ़኶ଷпክᆎᆎφ኶Ȃ൑ੂᘁ२࣐ঐրึ‫ي‬ਣ ෉ϟϊ़ᖃ२ଷпੂ኶ூ‫ژ‬ϟ҂ְᘁ२Ȅʳ. A. B. C. 79. ๗! ! ‫ݏ‬ ஆҐᢅ᜹ପРஊᎵஆϟਞ‫ݏ‬ Ս߇௳લ 120 ЉࡤϟѯᢋҪІ‫ݏ‬ುϑ֖༂ ዤ‫ޒ‬ᄙȂӶ‫ݏ‬ು੒ࢴࡤ஡ᆎφ௦ᆎܼϛӤᢅ᜹ ପРஊᎵஆϜȂஊᎵ 8 Љࡤ֊Ѡᢏᄇ‫ژ‬ᆎφึ ߄ȇክᆎࡤ 20 ЉȂϛӤପРஊᎵஆαϟᆎφึ ߄౦Ѡႁ 100%Ȃԫਣᆎφ़ϟಒΚбဩϑߞ яȇӶஊᎵ 50 ЉࡤѠُಒήбဩߞяȂ‫ٯ‬գᓎ ຀਴ߞяȄӶ 5 ᆎᢅ᜹ପРஊᎵஆϜȂп 1/2 MS ஊᎵஆᄈߵໍϊ़‫يึޠ‬ਞ‫ݏ‬ഷ‫ٺ‬Ȃϊ़ึ‫ي‬ ໧ࢳՎಒ 4 ‫ڸ‬ಒ 5 ਣ෉‫ޠ‬Ӭ‫ڂ‬Щ‫پ‬Ӕႁ 39.1%. D. E. შ 1ˁʳ ѯᢋҪІᆎφ़ϟϛӤึ‫ي‬ਣ෉ȄʻA) ਣ෉ 1Ȃ঩౩ᡞȇ(B) ਣ෉ 2ȂߞяಒΚбဩȇ(C) ਣ෉ 3ȂಒΚ бဩԚዤߞяಒΡбဩȇ(D) ਣ෉ 4ȂಒΡбဩτܼ 0.5 cmȇ(E) ਣ෉ 5ȂಒΡбဩԚዤಒήбဩτܼ 0.5 cm (Bar = 5 mm)Ȅʳ Fig. 1. Stages of seedling developmental of Bletilla formosana in in vitro culture. (A) Stage 1, protocorm; (B) Stage 2, protocorm with 1st leaf; (C) Stage 3, formation of 2nd leafʳ<ʳ0.5 cm in length; (D) Stage 4, formation of 2nd leafʳ> 0.5 cm in length; (E) Stage 5, formation of 3rd leafʳ> 0.5 cm in length. (Bar = 5 mm).

(4) ѯᢋၼཿः‫ف‬. 80. ಒ 60 ‫ڣ‬. (ߓ 1)Ȃᇅ‫ڐ‬у೏౪֖ᡘ຀ৰ౵ȂՅп VW ஊ ᎵஆഷৰȄЩၷᆎφ़ϟ൑ੂᘁ२ȂөପРஊ ᎵஆϜࠍп MS ஊᎵஆഷ‫ٺ‬Ȃ‫ڐ‬ಒ 4 Іಒ 5 ึ ‫෉ي‬ϟ൑ੂᘁ२ϸր࣐ 12.2 mg ᇅ 20.2 mgȂ‫ڐ‬ Ԫ࣐ 1/2 MS ஊᎵஆϟ 10.7 mg ᇅ 12.7 mgȄ. 0.1%ࣁ‫ܓ‬ᆈ఻ђᄈѯᢋҪІึ߄І़෉ ϟਞ‫ݏ‬ ܼϛӤᢅ᜹ପРϟஊᎵஆϜ఻ђ 0.1%ࣁ ‫ܓ‬ᆈȂӶஊᎵ 50 ЉࡤөପРஊᎵஆϜϟᆎφึ. ಒ2෉. ߄౦ࣲႁ 100%Ȃᡘұᎍ໕ࣁ‫ܓ‬ᆈ‫఻ޠ‬ђϛཽ ኈ៫ᆎφ‫߄ึޠ‬Ȅϊ़‫يึޠ‬Ӥኻп 1/2 MS ஊᎵஆ࣐ഷ‫ٺ‬Ȃಒ 4 ‫ڸ‬ಒ 5 ึ‫ي‬ਣ෉‫ޠ‬ϊ़Ӭ ‫ڂ‬Щ‫ ࣐پ‬46.2%ȇMS ᢅ᜹ஊᎵஆ‫ڐ‬ԪȂӬ‫ڂ‬Щ ‫پ‬Ҽႁ 38.6% (ߓ 2)Ȅп൑ੂᘁ२‫ࣽٿ‬Ȃ1/2 MS ஊᎵஆᇅ MS ஊᎵஆϜȂಒ 5 ึ‫ي‬ਣ෉ϟϊ़ ൑ੂᘁ२ϸր࣐ 20.0 mg І 20.8ʳ mg Ӥӗഷ ାȂՅп VW ஊᎵஆϟ 7.4 mg ഷৰȄᇅґ఻ђ ࣁ‫ܓ‬ᆈϟࣻӤପРஊᎵஆࣻЩ (ߓ 1)Ȃᗷดึ. ߓ 1. ϛӤᢅ᜹ପРஊᎵஆᄈѯᢋҪІณຄክᆎᆎφึ߄ᇅϊ़ึ‫ي‬ϟኈ៫ʳ Table 1. Effect of different basal salt media on in vitro seed germination and seedling development of Bletilla formosana Seed Basal salt germination formula z (%) y HYPONeX. Fresh weight of seedling at various developmental stages (mg/seedling). % of seedling at various developmental stages x 1+2. 3. 4+5. 100 a w 66.5 ± 2.21 b 16.3 ± 1.87 b 17.2 ± 0.63 b. 2. 3. 4. 5. 3.3 ± 0.14 bc. 5.5 ± 0.32 b. 7.8 ± 0.43 cd 10.5 ± 0.56 c. Kundson. 98 a. 67.4 ± 2.70 b. 9.2 ± 0.96 c 21.4 ± 1.85 b. 2.7 ± 0.13 cd. 4.3 ± 0.28 c. 8.5 ± 0.42 c. 10.2 ± 0.83 c. VW. 99 a. 90.4 ± 0.72 a. 1.8 ± 0.10 d. 4.3 ± 0.30 a. 4.8 ± 0.40 bc 7.0 ± 0.48 d. 10.2 ± 0.45 c. 6.8 ± 0.69 c. 1/2 MS. 100 a. 38.1 ± 1.57 c 22.9 ± 2.51 a 39.1 ± 3.60 a. 3.7 ± 0.24 ab. 7.7 ± 0.42 a 10.7 ± 0.51 b. 12.7 ± 0.46 b. MS. 100 a. 72.0 ± 4.84 b 12.5 ± 2.67 bc15.5 ± 2.24 b. 2.4 ± 0.17 d. 7.0 ± 0.30 a 12.2 ± 0.28 a. 20.2 ± 0.38 a. z. y x w. Bacto agar (0.8%) and sucrose (3%) were used to prepare basal salt media, using the formula of HYPONeX 7-6-19 (HYPONEX, USA), Kundson (Duchefa, Netherlands), VW (Duchefa, Netherlands), 1/2 MS (Duchefa, Netherlands), or MS (Duchefa, Netherlands). Seed germination was recorded at 50 days after sowing. Refer to Figure 1 for development stages of plants. Means ± standard error (n = 5). Means within each column followed by with the same letter are not significantly different at 5% level by LSD test.. ߓ 2. ֥ԥ 0.1%ࣁ‫ܓ‬ᆈϟϛӤᢅ᜹ପРஊᎵஆᄈณຄክᆎϟѯᢋҪІᆎφึ߄ᇅϊ़ึ‫ي‬ϟኈ៫ Table 2. Effect of basal salt media amended with 0.1% charcoal on in vitro seed germination and seedling development of Bletilla formosana Seed Basal salt germination formula z. (%) y w. Fresh weight of seedling at various developmental stages (mg/seedling). % of seedling at various developmental stages x 1+2. 4+5. 2. 3. 4. 5. HYPONeX. 100 a. 0.7 ± 0.27 b 29.6 ± 3.69 c. 1.8 ± 0.14 c. 3.6 ± 0.26 c. 7.4 ± 0.43 c. 14.7 ± 0.45 b. Kundson. 100 a. 61.2 ± 3.04 b 20.3 ± 1.95 a 18.5 ± 1.31 d. 3.4 ± 0.17 b. 7.1 ± 0.27 b. 11.4 ± 0.43 a. 13.8 ± 0.52 b. VW. 100 a. 79.7 ± 2.08 a. 4.1 ± 0.60 b 16.2 ± 1.64 d. 2.8 ± 0.14 b. 3.4 ± 0.19 c. 3.8 ± 0.26 d. 7.4 ± 0.20 c. 1/2 MS. 100 a. 29.9 ± 3.98 c 23.9 ± 1.67 a 46.2 ± 2.33 a. 5.9 ± 0.46 a. 6.8 ± 0.35 b. 9.9 ± 0.31 b. 20.0 ± 0.72 a. MS. 100 a. 38.9 ± 1.25 c 22.3 ± 2.10 a 38.6 ± 1.23 b. 5.8 ± 0.33 a. 9.2 ± 0.51 a. 10.5 ± 0.56ab. 20.8 ± 0.51 a. z. y x w. 69.7 ± 3.65 b. 3. Bacto agar (0.8%) and sucrose (3%) were used to prepare basal salt media, using the formula of HYPONeX 7-6-19 (HYPONEX, USA), Kundson (Duchefa, Netherlands), VW (Duchefa, Netherlands), 1/2 MS (Duchefa, Netherlands), or MS (Duchefa, Netherlands). Seed germination was recorded at 50 days after sowing. Refer to Figure 1 for development stages of plants. Means ± standard error (n = 5). Means within each column followed by with the same letter are not significantly different at 5% level by LSD test..

(5) ѯᢋҪІณຄክᆎ!. ߄౦ࣻծȂկ఻ђࣁ‫ܓ‬ᆈԥֆܼϊ़ϟϸϾȂ ܼಒ 4 Іಒ 5 ึ‫෉ي‬ϟϊ़ԥၷା‫ޠ‬Щ‫پ‬ȄӶ ᘁ२Р८Ȃпಒ 5 ึ‫ࣽٿ෉ي‬Ȃଷ VW І MS ஊᎵஆѵȂөᢅ᜹ପРӶ఻ђ 0.1%ࣁ‫ܓ‬ᆈࡤϟ ൑ੂᘁ२ࣲԥቩђȄʳ. ϛӤᐩ࡚ϟॸᒃԥᐡ఻ђ‫ޑ‬ପӬ఻ђ‫ܗ‬ϛ ఻ђ 0.1%ࣁ‫ܓ‬ᆈϟਞ‫ݏ‬ Ҧܼ 1/2 MS ᢅ᜹ஊᎵஆᄈѯᢋҪІϊ़ ึ‫ي‬ϟߵໍਞ‫ݏ‬ၷ‫ڐ‬уᢅ᜹ପР࣐‫ ߓ( ٺ‬1)Ȃ ӱԫп 1/2 MS ᢅ᜹ପР࣐ஆҐஊᎵஆȂଷ఻ђ 2.5–10%ϟ 4 ᆎϛӤ֥໕‫ॸޠ‬ᒃѵȂ‫ٯ‬ӤਣЩ ၷ 0.1%ࣁ‫ܓ‬ᆈ఻ђᄈᆎφึ߄Іึ‫ي‬ϟኈ ៫Ȅ๗‫ݏ‬Ԅߓ 3 ‫ܛ‬ұȂӶஊᎵஆϜ఻ђॸᒃ‫ٯ‬ ϛཽኈ៫ᆎφึ߄Ȃᆎφึ߄౦ϬѠႁ 100%Ȅ ॸᒃ֥໕ᇅࣁ‫ܓ‬ᆈϟ఻ђᄈϊ़‫يึޠ‬ԇӶ྄ ᡘ຀һདձңȂկณ፤఻ђ‫ܗ‬ґ఻ђࣁ‫ܓ‬ᆈࣲ п֥ 2.5%ॸᒃϟஊᎵஆܼಒ 4 Іಒ 5 ึ‫ڏ෉ي‬ ԥၷାЩ‫پ‬ϟϊ़ (28.3%) (ߓ 3)Ȃկᇅґ఻ђ. 81. ॸᒃϟ 1/2 MS ஊᎵஆࣻၷ (ߓ 1ȃ2)Ȃ఻ђॸ ᒃࡤᄈϊ़ึ‫ي‬ϟߓ౫ၷৰ (ߓ 3)ȄՅஊᎵஆ ᓎ຀ॸᒃ֥໕‫ޠ‬ቩђȂᄈϊ़ึ‫ي‬ԥϛًኈ ៫Ȃ֥ 10%ାᐩ࡚ॸᒃϟஊᎵஆसґ఻ђ 0.1% ‫ܓࣁޠ‬ᆈȂϊ़ึ‫ي‬τӼ୅Ӷಒ 1 І 2 ึ‫ي‬ਣ ෉ (ߓ 3)Ȃйτഌϸϑึ߄ϟᆎφณ‫ߞݳ‬Ԛ‫׈‬ ᐍϟෛੂ (შ 2)Ȅॸᒃ֥໕ᇅࣁ‫ܓ‬ᆈ఻ђଷಒ 3 ึ‫ي‬ਣ෉ѵȂ‫ڐ‬уึ‫ي‬ਣ෉ᄈϊ़ᘁ२ࣲԇ Ӷ྄ᡘ຀һདձңȂஊᎵஆϜ఻ђॸᒃᗷดᄈ ϊ़‫ޠ‬ᘁ२ԥቩђ‫ޠ‬ձңȂկଷ 2.5%ॸᒃѵȂ ණାॸᒃᐩ࡚ϟቩ२ਞ‫҇ݏ‬໹ପӬࣁ‫ܓ‬ᆈ‫఻ޠ‬ ђ (ߓ 3)Ȅ. ϛӤᐩ࡚ϟ଼ႜᖧԥᐡ఻ђ‫ޑ‬ପӬ఻ђ‫ܗ‬ ϛ఻ђ 0.1%ࣁ‫ܓ‬ᆈϟਞ‫ݏ‬ ஡αख़ၑᡜп଼ႜᖧх෈ॸᒃ఻ђܼክᆎ ஊᎵஆϜȂ๗‫ݏ‬ᡘұ଼ႜᖧ‫఻ޠ‬ђϛཽኈ៫ᆎ φึ߄Ȃᆎφึ߄౦ࣲѠႁ 100%ȄϛӤᐩ࡚ ଼ႜᖧᇅ఻ђࣁ‫ܓ‬ᆈᄈѯᢋҪІᆎφึ߄ᇅϊ. ߓ 3ˁʳ 1/2 MS ஆҐᢅ᜹ஊᎵஆ఻ђ 0.1%ࣁ‫ܓ‬ᆈᇅϛӤ֥໕ॸᒃ‫ݬ‬ᄈณຄክᆎϟѯᢋҪІᆎφึ߄ᇅϊ़ึ‫ي‬ ϟኈ៫ʳ Table 3. Effect of charcoal and banana homogenates in 1/2 MS medium on in vitro seed germination and seedling development of Bletilla formosana Banana Seed Charcoal homogenate germination (%) z (%) z (%) y. % of seedling at various developmental stages x 1+2 3 4+5. Fresh weight of seedling at various developmental stages (mg/seedling) 2 3 4 5. 0.0. 2.5. 100 a. 36.9 ± 1.93 d 37.7 ± 1.69 b 25.4 ±1.05 a. 6.2 ± 0.40 a. 7.3 ± 0.45 b. 11.7 ± 0.22 cd 25.0 ± 0.25 c. 0.1. 2.5. 100 a. 40.7 ± 1.17 d 31.0 ± 2.60 c 28.3 ± 1.98 a. 5.9 ± 0.47 a. 9.2 ± 0.36 a. 10.8 ± 0.56 d. 0.0. 5.0. 100 a. 35.0 ± 1.64 d 45.7 ± 1.41 a 19.3 ± 1.66 b. 2.7 ± 0.17 de 3.3 ± 0.18 e. 7.8 ± 0.30 e. 14.5 ± 0.36 e. 0.1. 5.0. 100 a. 58.6 ± 3.09 c 23.7 ± 1.73 d 17.7 ± 1.72 b. 4.7 ± 0.23 b. 7.6 ± 0.38 b. 12.8 ± 0.40 c. 24.9 ± 0.41 c. 21.1 ± 0.45 d. 0.0. 7.5. 100 a. 70.4 ± 2.87 b 14.7 ± 1.48 e 14.9 ± 1.65 b. 2.4 ± 0.17 e. 4.3 ± 0.23 d. 6.2 ± 0.30 f. 13.6 ± 0.26 e. 0.1. 7.5. 100 a. 74.5 ± 0.95 b. 6.4 ± 0.56 f 19.1 ± 1.42 b. 3.4 ± 0.22 cd 9.1 ± 0.32 a. 92.1 ± 0.94 a. 5.9 ± 0.74 f. 5.5 ± 0.29 c. 17.3 ± 0.50 a Ё w. 26.0 ± 0.31 b Ё. 4.2 ± 0.17 bc 8.9 ± 0.37 a. 15.4 ± 0.44 b. 28.6 ± 0.53 a. 0.0. 10.0. 98 a. 0.1. 10.0. 100 a. 0.0 ± 0.00 c. 74.2 ± 1.83 b 11.2 ± 0.81 e 14.6 ± 1.12 b. 4.6 ± 0.21 b. Significance. z y x w v. Charcoal (C). NS v. **. **. NS. NS. **. **. **. Banana (B). NS. **. **. **. **. **. NS. NS. C×B. NS. **. **. **. **. NS. **. *. Charcoal and banana homogenates were added to the medium containing 0.8% bacto agar, 3% sucrose and 1/2 MS (Duchefa, Netherlands). Seed germination was recorded at 50 days after sowing. Refer to Figure 1 for development stages of plants. No seedling. NS = Non-significant; *, **=significant at 0.05 and 0.01 levels, respectively..

(6) 82. ѯᢋၼཿः‫ف‬. ಒ 60 ‫ڣ‬. ಒ2෉. შ 2. ѯᢋҪІܼϛӤ֥໕ॸᒃϟ 1/2 MS ஆҐᢅ᜹ஊᎵஆʳ (A)ʳ ґ఻ђ‫ܗ‬ʳ (B) ఻ђ 0.1%ࣁ‫ܓ‬ᆈȂஊᎵ 50 Љϟ ᆎφึ߄ᇅϊ़ึ‫ي‬௒‫ם‬ȄშϜၑᆔҦѿՎѢॸᒃϟ֥໕ϸր࣐ 0%ȃ2.5%ȃ5.0%ȃ7.5%І 10.0%Ȅϊ़ܼ (B) ఻ђᇅ (A) ґ఻ђ 0.1%ࣁ‫ܓ‬ᆈஊᎵஆϜᚠՔԥ‫ܛ‬ৰ౵Ȅʳ Fig. 2. Seedlings of Bletilla formosana in 1/2 MS basal salt medium containing banana homogenates without (A) or with (B) 0.1% charcoal for 50 days. Left to right: Concentrations of banana homogenates in the medium, 0% (left tube), 2.5%, 5.0%, 7.5% and 10% (right tube). Note differences in color of seedlings between the treatments of 0.1% charcoal (B) and the treatments without charcoal (A).. ़ึ‫ࣲي‬ԇӶ྄ᡘ຀һདձң (ߓ 4)Ȃ‫ڐ‬Ϝп ֥ԥ 7.5%଼ႜᖧᇅ 0.1%ࣁ‫ܓ‬ᆈಣӬ‫ޠ‬ஊᎵஆ ࣐ഷ‫ٺ‬Ȃܼಒ 4 Іಒ 5 ึ‫ي‬ਣ෉‫ޠ‬ϊ़Щ‫پ‬ႁ 67.2%Ȃಒ 5 ึ‫ي‬ਣ෉‫ޠ‬ϊ़ϟ҂ְ൑ੂᘁ२ Ѡႁ 26 mgȇկस༊пᘁ२‫ࣽٿ‬Ȃࠍп 5.0%଼ ႜᖧ‫఻ٯ‬ђ 0.1%ࣁ‫ܓ‬ᆈϟஊᎵஆ‫ ޠ‬30.7 mg ࣐ ഷାȄөᐩ࡚ϟ଼ႜᖧପӬࣁ‫ܓ‬ᆈ఻ђᄈϊ़ ึ‫ي‬І२໕ְԥߵໍਞ‫ݏ‬Ȃկࣻᄈܼϛ଼֥ႜ ᖧІࣁ‫ܓ‬ᆈϟஊᎵஆ (ߓ 1)Ȃାᐩ଼࡚ႜᖧ. (7.5%ȃ10%) ஊᎵஆϟϊ़Ңߞϛ‫ښ׼ڨ‬Ȃࣲ ѠߞԚ‫׈‬ᐍෛੂ (შ 3)Ȅ. ଇ! ! ፤ ៍ऌෛ‫ޑ‬ϟᆎφߩளϊȂथϸϾѬႁ༬౩ थ໧ࢳ (globular embryo stage)Ȃϛ֥थ‫ٱ‬Ȃ‫ء‬ ԥφဩȃथ਴ȃथ߄๊ᎵϸᓾᙡᏣۣȂሰစҦ ѵӶϟᕘძ‫ٽ‬ᔗщ٘‫ޠ‬Ꮅϸ (៍ຄӔҢ‫ܗ‬ณຄ ክᆎஊᎵஆ) Р૗໸ւ຃ึߞτ (Arditti 1967;.

(7) ѯᢋҪІณຄክᆎ!. 83. 100%Ȃйϊ़໸ւึ‫ي‬ᇅҢߞȂѫпϑ໡ບϟ ‫ݏ‬ುໍ՘ክᆎȂҼԥًԂϟึ߄౦ (ၦਠґᡘ ұ)Ȃԫ๗‫ݏ‬ၷࣻծܼ Nagashima (1982) ϟः ‫ف‬ȂՅᇅ Lin et al. (1994) ‫ޠ‬๗‫ݏ‬ԥ‫ܛ‬ৰ౵ȄӶ ᆎφԚዤ࡚‫ޠ‬ኈ៫Р८ȂӶ Lin et al. (1994) ‫ޠ‬ ः‫ف‬Ϝࡿя༂ዤۧґ໡ບϟѯᢋҪІᆎφณ‫ݳ‬ ຃߄ȂҐၑᡜ‫ٻ‬ң༂ዤۧґ໡ບϟѯᢋҪІ‫ݏ‬ ುܼϛӤᢅ᜹ϟஊᎵஆໍ՘ณຄክᆎȂܼክᆎ ࡤ 8 Љ֊Ѡُᆎφึ߄Ȃ20 Љࡤ຃߄౦ႁ 100%Ȃᆎφ຃߄ᇅϊ़Ңߞഁ࡚ၷ Lin et al. (1994) ᇅ Yeh et al. (1995) ϟᢏᄇ‫ץ‬ഁȂѠ૗ ᇅ؆ਠ‫ྜٿ‬ᇅณຄክᆎႇโϛӤՅԥ‫ܛ‬ৰ౵Ȃ ӱԫȂӶҐၑᡜϜȂ௱ข௳લࡤ 120 Љϟѯᢋ ҪІ‫ݏ‬ುԚዤ࡚ᔗ၏ᎍӬܼณຄክᆎϟңȄԫ ѵȂ‫ש‬উւңࡊྤίϑ໡ບ 2 ໋‫ݏ‬ುϟᆎφໍ՘ ณຄክᆎȂึ౫‫ڐ‬ᆎφϬ‫ڏ‬ԥًԂϟึ߄૗Ω (ၦਠґᡘұ)Ȃ௱ขΚঐ‫ڏ‬ԥًԂ๗ᄃ౦ϟѯᢋ ҪІ‫ݏ‬ು‫߄ึڐ‬ᎍ෉ᔗԥࣻ࿌‫ޠ‬ਣ໣ጓ൝ȂӶϛ Ӥ‫߄ึޠ‬ஊᎵஆϜҼԥࣻ࿌‫ޠ‬ᎍᔗ‫ܓ‬Ȅ. Lee 1990)Ȅ៍ऌෛ‫ޑ‬ᆎφԚዤ࡚Ҽኈ៫‫ڐ‬ᆎφ ‫ޠ‬຃߄૗ΩȂϛӤ៍឵໣‫ڐ‬ഷ‫ݏٺ‬ು௵Ԟਣ໣ ԥ‫ܛ‬ৰ౵ȂӶፆፇ៍Ҫ߇ᚖһᆎ‫ޠ‬ः‫ف‬ᡘұȂ ௳લࡤ 110 Љ௵Ԟϟ‫ݏ‬ುӶክᆎࡤ 10 Љ֊Ѡึ ߄Ȃस‫۾‬Վ‫ݏ‬ು༂ዤࣦՎ໡ບϘ՘ክᆎȂҼϛ ኈ៫ᆎφ຃߄૗ΩІ঩౩ᡞึ‫( ي‬Tu & Lee 1987)ȇկӶ࿳х៍ (Mathews & Rao 1980)ȃൊ ඾Ꭿ៍ Cypripedium (Harvais 1980)ȃѯᢋҪІ (Lin et al. 1994) ๊៍឵‫ޠ‬ः‫ࠍف‬ᡘұȂܼ༂ዤ ෉௵Ԟϟ‫ݏ‬ುክᆎࡤȂᆎφϛึ߄‫߄ึܗ‬౦ϛ ‫ٺ‬ȂࣦՎኈ៫ϊ़ϟҢߞȂѠ૗঩ӱ࣐ᆎφԚ ዤࡤ‫ޑښ׼‬፵ϟ‫ם‬ԚȂໍՅኈ៫ᆎφึ߄ᇅҢ ߞȄӶҪІ឵‫ޠ‬ः‫ف‬ᡘұȂ‫ڐ‬ᆎφӶथ௦ߗԚ ዤਣክᆎึ߄౦ഷା (Nagashima 1982)ȇLin et al. (1994) ࠍᇰ࣐ѯᢋҪІпึ‫ي‬ՎനႫ෉ϟ ᆎφ຃߄౦ᇅϊ़Ңߞ࣐ഷ‫ٺ‬ȂՅ༂ዤ‫ݏ‬ುϟ ᆎφࠍณ‫߄ึݳ‬ȄӶҐၑᡜϜ௵ң௳લ 120 Љ ۧґ໡ບϟ༂ዤѯᢋҪІ‫ݏ‬ುክᆎȂӶөᆎϛ ӤପРஆҐᢅ᜹ϟஊᎵஆϜȂ‫߄ึڐ‬౦ࣲѠႁ. ߓ 4ˁʳ1/2 MS ஆҐᢅ᜹ஊᎵஆ఻ђ 0.1%ࣁ‫ܓ‬ᆈᇅϛӤ֥໕଼ႜᖧ‫ݬ‬ᄈѯᢋҪІᆎφึ߄ᇅϊ़ึ‫ي‬ϟኈ៫ Table 4.ʳEffect of charcoal and potato homogenates in 1/2 MS medium on in vitro seed germination and seedling development of Bletilla formosana Potato Seed % of seedling at various Charcoal homogenate germination developmental stages x (%) z (%) z (%) y 1+2 3 4+5 0.0 2.5 100 a 22.2 ± 1.18 c 35.6 ± 1.59 ab 42.2 ± 1.21 e. Fresh weight of seedling at various developmental stages (mg/seedling) 2 3 4 5 5.1 ± 0.23 b 4.2 ± 0.33 d 6.2 ± 0.31 e 11.0 ± 0.30 e. 0.1. 2.5. 100 a. 26.0 ± 1.36 c 20.4 ± 0.77 d 53.7 ± 2.01 c. 5.8 ± 0.17 a 11.5 ± 0.25 a 12.8 ± 0.36 c 24.7 ± 0.85 b. 0.0. 5.0. 100 a. 13.8 ± 1.00 d 26.0 ± 1.44 d 60.2 ± 2.40 b. 6.3 ± 0.23 a 6.6 ± 0.43 c 10.1 ± 0.51 d 15.1 ± 0.29 d. 0.1. 5.0. 100 a. 10.1 ± 1.11 d 39.6 ± 2.47 a 50.3 ± 2.39 cd. 4.1 ± 0.34 c 7.1 ± 0.45 c 15.1 ± 0.67 b 30.7 ± 0.56 a. 0.0. 7.5. 100 a. 61.1 ± 0.54 a 11.4 ± 0.59 e 27.5 ± 0.70 f. 2.7 ± 0.09 d 4.3 ± 0.33 d. 0.1. 7.5. 100 a. 0.0 ± 0.00 e 32.8 ± 2.13 c 67.2 ± 2.13 a. -w. 6.9 ± 0.37 e 14.9 ± 0.36 d. 10.3 ± 0.52 b 16.3 ± 0.33 a 26.1 ± 0.63 b. 0.0. 10.0. 100 a. 57.6 ± 2.82 a 18.9 ± 1.19 d 23.5 ± 2.76 f. 2.5 ± 0.17 d 4.5 ± 0.21 d. 0.1. 10.0. 100 a. 44.1 ± 1.87 b 12.0 ± 0.71 e 43.9 ± 2.44 de. 4.0 ± 0.34 c 6.7 ± 0.41 c 13.1 ± 0.42 c 25.5 ± 0.34 b. 9.5 ± 0.29 d 17.5 ± 0.24 c. Significance. z y x w v. Charcoal (C). NS v. **. **. **. **. **. **. **. Potato (P). NS. **. **. **. **. **. **. **. C×P. NS. **. **. **. **. **. **. **. Charcoal and potato homogenates were added to the medium containing 0.8% bacto agar, 3% sucrose and 1/2 MS (Duchefa, Netherlands). Seed germination was recorded at 50 days after sowing. Refer to Figure 1 for development stages of plants. No seedling. NS = Non-significant; *, ** = significant at 0.05 and 0.01 levels, respectively..

(8) 84. ѯᢋၼཿः‫ف‬. ಒ 60 ‫ڣ‬. ಒ2෉. შ 3. ѯᢋҪІܼϛӤ֥໕଼ႜᖧϟ 1/2 MS ஆҐᢅ᜹ஊᎵஆ (A)ʳ ґ఻ђ‫ܗ‬ʳ (B) ఻ђ 0.1%ࣁ‫ܓ‬ᆈஊᎵ 50 Љϟ ᆎφึ߄ᇅϊ़ึ‫ي‬௒‫ם‬ȄშϜၑᆔҦѿՎѢ଼ႜᖧϟ֥໕ϸր࣐ 2.5%ȃ5.0%ȃ7.5%І 10.0%Ƕϊ़ܼ (B) ఻ ђᇅ (A) ґ఻ђ 0.1%ࣁ‫ܓ‬ᆈஊᎵஆϜᚠՔԥ‫ܛ‬ৰ౵Ȅʳ Fig. 3. Seedlings of Bletilla formosana in 1/2 MS basal salt medium containing potato homogenates without (A) or with (B) 0.1% charcoal for 50 days. Left to right: Concentrations of potato homogenates in the medium, 0% (left tube), 2.5%, 5.0%, 7.5% and 10% (right tube). Note differences in color of seedlings between the treatments of 0.1% charcoal (B) and the treatments without charcoal (A).. MS ஊ Ꮅ ஆ ପ Р ϟ ᖃ ᚕ φ ᐩ ࡚ ࣐ 93.8 mMȂ1/2 MS ஊᎵஆϟᖃᚕφᐩ࡚࣐ 46.9 mM (Tu & Lee 1988)ȂKundson (Knudson 1946) ᇅ VW (Vacin & Went 1949) ପРᖃᚕφᐩ࡚ङ࣐ 30 mM (Lo et al. 2008)Ȃ߇᝙ 1 ဵ (HYPONeX 7-6-19) ࣐୧ཿҢ౱ϟፓӬ޴ਠȂ‫ڐ‬ᖃ N%ၷα ख़ᢅ᜹ஊᎵஆְմ (Tu & Lee 1988)ȄLin et al.. (1994) ‫ޠ‬ः‫ࡿف‬яȂMS ஆҐᢅ᜹Іጶᑦ࢑ѯ ᢋҪІ঩౩ᡞึ‫ي‬Ԛϊ़ϟ्҇ᕋᎵԚϸȂ2 ॼ໕ϟ MS ஆҐᢅ᜹ࠍཽ౱Ң།ড় (ᖃᚕφᐩ ࡚ႁ 187.6 mM)Ȃ‫ٻ‬ϊ़ߓҫዺᕼᡑ‫ם‬й‫ښ׼‬ ҢߞȄ਴ᐄ Ichihashi (1979) ‫ޠ‬ः‫ف‬ᡘұȂᇰ࣐ ᖃᚕφᐩ࡚Ӷ 60 mM ਣၷԥւܼҪІӵαഌ ҢߞȄӶҐः‫ف‬ϜѯᢋҪІܼ MSȃ1/2 MSȃ.

(9) ѯᢋҪІณຄክᆎ!. KundsonȃVW І߇᝙ 1 ဵᢅ᜹ପРஊᎵஆϜȂ ᆎφึ߄౦ࣲѠႁ 100%Ȃйϊ़Ѡ໸ւึ‫ي‬Ȃ ϥᆎᢅ᜹ପРஊᎵஆϜп 1/2 MS ϊ़ึ‫࣐ي‬ ഷ‫ ߓ( ٺ‬1)ȂӱԫȂӶҪІ឵ࣻၷܼ‫ڐ‬у៍ऌ ෛ‫ޑ‬Ȃᆎφึ߄ᇅϊ़ึ‫ي‬ᄈᢅ᜹ᐩ࡚ϟᎍᔗ ၷኅȂ७մᢅ᜹ᐩ࡚ᔗԥւܼᆎφึ߄ᇅϊ़ Ңߞ (Arditti 1967; Ichihashi 1979; Tu & Lee 1988; Lu & Lee 1990; Wu & Lee 1991)Ȅ Ӷ៍ऌෛ‫ޠޑ‬ณຄክᆎІᚕᡞஊᎵϜள఻ ђࣁ‫ܓ‬ᆈȂӱ‫ڏڐ‬ԥ֝ߤ⍈᜹ϾӬ‫ޑ‬ȃ‫ښ׼‬ႇ ੋϾሖષࣁ‫ܓ‬ȃ७մፒϾ౦ȃߵໍ߄ȃ਴ᇅෛ ੂ Ң ߞ ȃ ࠯ ᄙ ึ Ң ๊ ፓ ᚖ ϟ ѓ ਞ (Tisserate 1979; Patel & Thorpe 1984; Pan & Staden 1998)ȂӶҐၑᡜϜȂܼϛӤᢅ᜹ପРஊᎵஆϜ ఻ђ 0.1%‫ܓࣁޠ‬ᆈȂᄈᆎφ‫ء߄ึޠ‬ԥ‫ٯ‬ณϛ ًϟኈ៫Ȃйࣁ‫ܓ‬ᆈ‫఻ޠ‬ђѠߵໍϊ़ึ‫ي‬ᇅ Ңߞ (ߓ 2–4 Іშ 2)ȄErnst (1975) ‫ޠ‬ः‫ف‬Ҽᡘ ұȂࣁ‫ܓ‬ᆈ‫఻ޠ‬ђᄈ៍ऌෛ‫ءޑ‬ԥϛًኈ៫Ȃ կ Van (1987) ᄈ ܼ ঈ ຾ ៍ ឵ Anacamptis pyramidalis ๊ 18 ᆎዊࢹ៍ऌෛ‫ޠޑ‬ः‫ࠔف‬ᡘ ұȂࣁ‫ܓ‬ᆈ‫఻ޠ‬ђཽ७մ‫ڐ‬ᆎφึ߄౦ᇅ‫ښ׼‬ ϊ़ึ‫ي‬ȂՅ Lin et al. (1994) ‫ޠ‬ः‫ࡿف‬я 0.1–0.3%ϟࣁ‫ܓ‬ᆈᎍӬѯᢋҪІᆎφึ߄ᇅҢ ߞȂկ 0.5%пαࣁ‫ܓ‬ᆈ‫఻ޠ‬ђࠍ७մᆎφึ߄ ౦ᇅ‫ښ׼‬ϊ़Ңߞᇅึ‫ي‬ȄӶҐၑᡜϜȂп 0.1%ࣁ‫ܓ‬ᆈ఻ђܼϛӤᢅ᜹ପРஊᎵஆϜȂ๗ ‫ݏ‬ᡘұϊ़Ңߞၷ࣐‫ץ‬ഁȂ൑ੂᘁ२Ҽԥቩ ђȂᄈѯᢋҪІณຄክᆎϊ़ϟҢߞึ‫ڏي‬ԥ ߵໍϟਞ‫ݏ‬Ȃկ‫ڨ‬ԥᐡ‫ޑ‬ኈ៫Ȃ೼ΚᘉӶॸᒃȃ ଼ႜᖧ఻ђၑᡜϜȂࣁ‫ܓ‬ᆈᇅ೼Ρᆎԥᐡ఻ђ ‫ޑ‬໣ְԥᡘ຀ϟһདਞᔗѠпӠᜍȄ Ӷ៍߇‫ޠ‬ಣᙒஊᎵϜȂஊᎵஆଷΠஆҐ‫ޠ‬ ᢅ᜹ϟѵȂளѫ՘఻ђጺ‫ݏ‬ҙߵໍϊ़ϟึ‫ي‬ (Arditti 1967)Ȃ‫ڐ‬Ϝॸᒃᇅ଼ႜᖧ࣐ၷளңϟ ጺ‫఻ݏ‬ђ‫ޑ‬ȂӶංᆎ៍ऌෛ‫ޠޑ‬ः‫ف‬ᡘұȂܼ ஊᎵஆϜ఻ђᎍ໕‫ॸޠ‬ᒃ (6–10%) ‫଼ܗ‬ႜᖧ (2–4%)Ȃᄈܼϊ़ึ‫ي‬ȃҢߞᇅᘁ२‫ޠ‬ණЁഎ. 85. ‫ڏ‬ԥًԂਞ‫( ݏ‬Lo et al. 2004; Shiau et al. 2005; Yan et al. 2006)ȇӶѯᢋΚဩ៍‫ޠ‬ः‫ف‬ҼᡘұȂ ஊᎵஆϜॸᒃ֥໕ႁ 150 g·L-ˬਣȂᆎφึ߄౦ ७մ (Juang & Lee 1986)ȄӶҐၑᡜϜ‫ٻ‬ң 2.5–10%ॸᒃ (ߓ 3) ᇅ଼ႜᖧ (ߓ 4) ‫఻ޠ‬ђ ‫ٯ‬ϛཽኈ៫ѯᢋҪІᆎφ‫߄ึޠ‬Ȃկॸᒃ‫֥ޠ‬ ໕ᡘ຀ኈ៫ϊ़ϟҢߞᇅึ‫ ߓ( ي‬3)Ȃ‫ٻ‬ң໕ ႁ 10%ਣȂᄈϊ़Ңߞཽ౱Ң‫ښ׼‬ձңȄկ଼ ႜᖧ‫఻ޠ‬ђᄈϊ़‫ޠ‬Ңߞᇅึ‫ࠍي‬ԥߵໍϟਞ ‫ ߓ( ݏ‬4)Ȃ‫ڐ‬Ϝп֥ԥ 7.5%଼ႜᖧᇅ 0.1%ࣁ‫ܓ‬ ᆈϟஊᎵஆᄈѯᢋҪІϟϊ़Ңߞߵໍਞ‫ݏ‬ഷ ‫ٺ‬ȂӶஊᎵ 50 ЉࡤȂಒ 4 Іಒ 5 ึ‫෉ي‬ϊ़౦ႁ 67.2%Ȃึ‫ ي‬5 ෉ϟϊ़҂ְᘁ२ႁ‫ ژ‬26.1 mgȄ ܼ Lin et al. (1994) ᇅ Yeh et al. (1995) ᄈѯᢋ ҪІณຄክᆎ‫ޠ‬ः‫ف‬ϜଷΠໍ՘ϛӤ MS ᢅ ᜹ȃጶᑦȃࣁ‫ܓ‬ᆈᐩ࡚ȃ‫ݏ‬ುԚዤ࡚ϟ೏౪ѵȂ ‫ٯ‬ґໍ՘ጺ‫఻ݏ‬ђ‫ޑ‬ᄈѯᢋҪІᆎφึ߄ᇅϊ ़Ңߞึ‫ي‬ኈ៫ϟ௥ଇȂҐः‫ف‬ϜѯᢋҪІᆎ φึ߄ᇅϊ़Ңߞࣲၷαख़Ᏹ޲ः‫ف‬๗‫( ݏ‬ஊ Ꮅ 135 Љ) ‫ץޠ‬ഁȂଷᇅᆎφϟԚዤ࡚ԥᜱ ѵȂᔗᇅክᆎஊᎵஆ‫ޠ‬ᢅ᜹ᐩ࡚пІᎍ࿌ϟԥ ᐡ఻ђ‫ໍߵޑ‬ϊ़ึ‫ي‬ԥᜱȄ Ґः‫ف‬ᙥҦϑ‫ޤ‬ஊᎵஆପР‫ޠ‬ᑣᒶȂପӬ ᎍ࿌ᐩ࡚ϟጺ‫఻ݏ‬ђ‫ޑ‬Іࣁ‫ܓ‬ᆈ఻ђȂւңܼ ѯᢋҪІϟณຄክᆎȂϛկᆎφϟึ߄౦ାȂ й૗ђഁϊ़ϟึ‫ي‬ᇅණା౮़ϟࠣ፵ȂණЁ ΠѯᢋҪІณຄክᆎᖆ෦ᡞ‫ق‬ϟਞ౦Ȃԫᖆ෦ ‫ق‬ಜѠණ‫ٽ‬τ໕ᓻًϟѯᢋҪІᆎ़ձ࣐Ң౱ ᛿؆‫࣐ܗ‬༫᛻ၦྜϟ‫ܛ‬ሰȄ. ЖңНᝧ (Literature cited) Arditti, J. 1967. Factors affecting the germination of orchid seeds. Bot. Rev. 33:1–97. Duan, J. X. and S. Yazawa. 1995. Floral induction and development in Phalaenopsis in vitro. Plant Cell Tiss. Organ Cult. 43:71–74. Ernst, R. 1975. Studies in asymbiotic culture of orchid. Amer. Orchid Soc. Cult. 44:12–18. Fu, Z. H., J. X. Zhang, H. L. Li, and B. Yang. 2006. Study.

(10) 86. ѯᢋၼཿः‫ف‬. on seed germination and rapid propagation of Bletilla striata Rchb. f. J. Wuhan. Bot. Res. 24:80– 82. (in Chinese with English abstract) Harvais, G. 1980. Scientific notes on a Cypripedium reqina of northwestern Ontario, Canada. Amer. Orchid Soc. Bull. 49:237–244. Ichihashi, S. 1979. Study on the media for orchid seed germination. III. The effect of total ionic concentration, cation/anion ratio, NH4+/NO3- ratio, and minor elements on the growth of Bletilla striata. New Phytol. 83:121–128. Juang, J. H. and N. Lee. 1986. Effect of niacin, coconut milk and banana homogenate on seed germination and seedling growth of Pleione formosana. J. Chinese Soc. Hort. Sci. 32:132–138. (in Chinese with English abstract) Knudson, L. 1946. A new nutrient solution for orchid seed germination. Amer. Orchid Soc. Bull. 15:214– 217. Lee. 1990. Embryo culture of orchids. J. Chinese Soc. Hort. Sci. 36:223–244. (in Chinese with English abstract) Lin, Y. J., C. C. Chuan, F. T. Yeh, N. Y. Chiu, and H. S. Tsay. 1994. Tissue culture of Bletilla formosana I. The influence of seed maturity and pretreament on seed germination and seeding development. J. Agric. Res. China 43:40–50. (in Chinese with English abstract) Lo, S. F., S. M. Nalawade, C. L. Kuo, C. L. Chen, and H S. Tsay. 2004. Asymbiotic germination of immature seeds, plantlet development and ex vitro establishment of plants of Dendrobium tosaense makino-A medicinally important orchid. In Vitro Cell Dev. Biol. Plant 40:528–535. Lo, S. F., C. L. Kuo, C. L. Chen, and H. S. Tsay. 2008. In vitro seed germination and mass propagation of Dendrobium moniliforme, a native species in Taiwan. J. Taiwan Agric. Res. 57:295–304. (in Chinese with English abstract) Lu, I. L. and N. Lee. 1990. In vitro germination of Cymbidium Dayanum. J. Chinese Soc. Hort. Sci. 36:198–209. (in Chinese with English abstract) Mathews, V. H. and P. S. Rao. 1980. In vitro multiplication of Vanda hybrids through tissue culture technique. Plant Sci. Lett. 17:383–389. Murashige, T. and F. Skoog. 1962. A revised medium for rapid growth and bioassays with tobacco tissue. ಒ 60 ‫ڣ‬. ಒ2෉. culture. Physiol. Plant. 15:473–491. Na, C., W. S. Kan, N. Y. Chiu, and R. J. Yang. 1978. Pharmacognostical researches on Bletillae tuber Pai-Chi. China Med. Coll. Annu. Bull. 9:454–467. (in Chinese with English abstract) Nagashima, T. 1982. Studies on the seed germination and embryogenesis in the Bletilla striata Rchb. f. and Calanthe discolor Lindl. J. Jpn. Soc. Hort. Sci. 51:82–93. (in Chinese with English abstract) Pan, M. J. and J. V. Staden. 1998. The use of charcoal in in vitro culture-A review. Plant Growth Regul. 26: 155–163. Patel, K. R. and T. A. Thorpe. 1984. In vitro differentiation of plantlets from embryonic explants of lodgepole pine (Pinus contorta Doughl. ex Loud). Plant Cell Tiss. Organ Cult. 3:131–142. Peng, L., X. D. Liu, H. Liu, and X. W. Liu. 2004. Tissue culture and rapid propagation of Bletilla striata (Thunb.) Rchb. f. Chinese Wild Plant Resources 23:65. (in Chinese with English abstract) Shiau, Y. J., S. R. Yang, U. C. Chen, C. N. Hsia, and H. S. Tsay. Effect of Vegetable and fruit homogenates on seedling development of Dendrobium moniliforme (L.) sweet culture in vitro. J. Agric. Res. China 54:23–31. (in Chinese with English abstract) Tisserat, B. 1979. Propagation of data palm (Phoenix dactylifera L.) in vitro. J. Exp. Bot. 30:1275–1283. Tu, M. C. and N. Lee. 1987. Effect of pollination time and capsule maturity on seed germination in Phalaenopsis white hybrid. J. Chinese Soc. Hort. Sci. 33:190–200. (in Chinese with English abstract) Tu, M. C. and N. Lee. 1988. Effect of nitrogen, sucrose concentration and light intensity on seed germination and seedling growth in Phalaenopsis white hybrid. J. Chinese Soc. Hort. Sci. 34:293– 302. (in Chinese with English abstract) Vacin, E. F. and F. W. Went. 1949. Some pH changes in nutrient solutions. Bot. Gaz. 110:605–613. Van, W. J. 1987. Effect of activated charcoal on in vitro propagation of Western European orchid. Acta Hort. 212:131–138. Wu, S. C. and N. Lee. 1991. In vitro seed germination of Phaius tankervilliae. J. Chinese Soc. Hort. Sci. 37:183–188. (in Chinese with English abstract) Yan, N., H. Hu, J. L. Huang, K. Xu, H. Wang, and Z. K. Zhou. 2006. Micropropagation of Cypripedium flavum through multiple shoots of seedlings.

(11) ѯᢋҪІณຄክᆎ!. 87. derived from mature seeds. Plant Cell Tiss. Organ Cult. 84:113–117.. Agric. Res. China 5:353–363. (in Chinese with English abstract). Yang, J. H., Q. J. Meng, J. D. Li, and H. T. Wu. 2002. Rapid propagation of Bletilla striata Reich b. f. by tissue culture. Shandong Sci. 15:13–16. (in Chinese with English abstract). Zhu, Y. Q. and X. G. Wang. 1999. Fast propagation techniques of Bletilla ochracea by means of tissue cultures. J. Zhejiang For. Coll. 16:164–169. (in Chinese with English abstract). Yeh, F. T., Y. C. Lin, C. C. Chen, N. Y. Chiu, and H. S. Tsay. 1995. Tissue culture of Bletilla formosana II. The influence of medium components on imamture seed germination and seeding development. J.. Zhang, J. X., Z. H. Fu, H. L. Li, and B. Yang. 2005. Relationship between embryo growth and seeds sprouting of Bletilla striata. Subtrop. Plant Sci. 34:32–35. (in Chinese with English abstract).

(12) 88. ѯᢋၼཿः‫ف‬. ಒ 60 ‫ڣ‬. ಒ2෉. Effect of Basal Salts, Active Charcoal, and Vegetable/Fruit Homogenates on Seed Germination and Seedling Development Bletilla formosana (Hayata) Schltr in In Vitro Culture1 Chin-Yi Tsao2, Uei-Chern Chen2, and Chi-Ni Hsia2,3 Abstract Tsao, C. Y., U. C. Chen, and C. N. Hsia. 2010. Effect of basal salts, active charcoal, and vegetable/fruit homogenates on seed germination and seedling development Bletilla formosana (Hayata) Schltr in in vitro culture. J. Taiwan Agric. Res. 60:77–88.. This study was conduced to determine effect to basal salts, active charcoal and vegetable/fruit homogenates on seed germination and seedling development of Bletilla formosana (Hayata) Schltr in in vitro culture. Bacto agar (0.8%) and sucrose (3%) were used to prepare the five culture media, using reported basal formulations of Knudson (Knudson 1946), VW (Vacin & Went 1949), MS (Murashige & Skoog 1962), 1/2 MS, or HYPONeX 7-6-19. Results showed that seeds of B. formosana were all germinated on these basal salt media after incubation at (25 ± 1к) for 8 days. Among the five basal salt media tested, the medium containing 1/2 MS was the best treatment for development and growth of seedling of B. formosana. Adding 0.1% active charcoal to each of the five basal salt media had no significant (P > 0.05) effects on seed germination as all the seeds germinated on each medium after incubation at (25 ± 1к) for 50 days. Fresh weight of 50-day-old seedling was 20.0 mg/plant for the treatment of 1/2 MS + active carbon and 20.8 mg/plant for MS +ʳ active charcoal but it was only 7.4 mg/plant for the treatment of VW +ʳactive charcoal. Adding banana or potato homogenates (at 2.5, 5.0, 7.5, 10%) to the medium containing 1/2 MS, with or without 0.1% active charcoal, had no negative effect on seed germination. Although both banana and potato substrates in the medium caused an increase in fresh weight of seedlings, the potato substrate was superior to the banana substrate for seedling development. This study suggests that the medium containing 1/2 MS salts, 0.1% active charcoal and 7.5% potato substrate is the best for seed germination and seedling development of B. formosana under aseptic conditions. Key words: Bletilla formosana, Seed germination, Seedling development, In vitro culture.. 1. Contribution No. 2502 from Taiwan Agricultural Research Institute (TARI), Council of Agriculture. Accepted: March 31, 2011. 2. Respectively, Assistant Researcher, Assistant Researcher, and Associate Researcher, Biotechnology Division, TARI, Taichung, Taiwan, ROC. 3. Corresponding author, e-mail: [email protected]; Fax: (04)23302806..

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