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Á¡µ³Á¤¨È—¥µŠ¡µ¦µµ¥¡´œ›»r RRIM600 ž¦³¤µ– 50 Á¤¨È—Ĝž¨Š œµ— 2 x 2 m 2 ¦—œÊεª´œ¨³ 1 ‡¦´ÊŠ Áª¨µ
ž¦³¤µ– 08.00 œ. Á¤ºÉ°˜oœ„¨oµ¥µŠ¤¸°µ¥»ž¦³¤µ– 3 Á—º°œ ‹¹ŠÁ„È ˜´ª°¥nµŠÁª¨µ 10.00 œ. ŞÁ˜¦¸¥¤ total RNA ‹µ„
ÁœºÊ °Á¥ºÉ°Ä „oµœÄ ¨Îµ˜oœ ¨³¦µ„ ˜µ¤ª·›¸ „µ¦ °ŠÁž¨ºÊ°Š (2549) ¨³ª·Á‡¦µ³®r„µ¦Â—Š°°„ °Š¥¸œ—o ª¥Áš‡œ·‡
sqRT-PCR
Ÿ¨ °ŠÃŽÁ—¸¥¤‡¨°Å¦—r¨³Ã¡¨¸Á°›¸¨¸œÅ„¨‡°¨˜n°„µ¦Â—Š°°„ TDFREP241 Ĝ˜oœ„¨oµ¥µŠ¡µ¦µ
Á¡µ³Á¨¸Ê¥ŠÁ°È ¤¦·Ã°¥µŠ¡µ¦µµ¥¡´œ›»r RRIM600 œ°µ®µ¦Â Ȋ¼˜¦ MS (Murashige and Skoog, 1962)
Á¤ºÉ °Á°È¤¦·Ã°Á‹¦·Áž} œ˜oœ„¨o µ¤¸ÄÂšo‡¼nš¸É 2 ‹¹ ŠœÎµÅžÄoĜ„µ¦š—¨°ŠÃ—¥ªµŠÂŸœ„µ¦š—¨°ŠÂ»n¤˜¨°—
(completely randomized design, CRD) „µ¦š—¨°Š¤¸ ‹Îµœªœ 2 „µ¦š—¨°Š ‡º° „µ¦š—¨°Š»—š¸É 1 ¤¸ š´ÊŠ®¤— 4 š¦¸˜-
Á¤œ˜r š¦¸ ˜Á¤œ˜r¨³ 5 ŽÊε ˜n¨³š¦¸˜Á¤œ˜rÁ˜·¤°µ®µ¦Á®¨ª¼˜¦ MS š¸É¤¸ n ªœŸ¤ÃŽÁ—¸¥¤‡¨°Å¦—r 0.5 䨵¦r ¨³
œÎ µÅžªµŠÁ¨¸Ê¥ŠÃ—¥®¤»œÁ®ª¸É¥Š—oª¥Â¦ŠÁ®ª¸É ¥Š 60 ¦°/œµš¸ £µ¥Ä˜o‡ªµ¤Á o¤ °ŠÂŠ 1,000 ¨´„¬r Áž}œÁª¨µ˜nµŠÇ „´ œ
‡º° š¦¸˜Á¤œ˜rš¸É 1, 2, 3 ¨³ 4 ªµŠÅªoÁž}œÁª¨µ 0, 0.5, 6 ´ÉªÃ¤ŠÂ¨³ 24 ´ÉªÃ¤Š ˜µ¤¨Îµ—´ „µ¦š—¨°Š»—š¸É 2 ¤¸š´Ê Š®¤—
4 š¦¸˜Á¤œ˜r š¦¸˜Á¤œ˜r ¨³ 5 ŽÊε ˜n¨³š¦¸˜Á¤œ˜r Á˜·¤°µ®µ¦Á®¨ª¼˜¦ MS š¸É¤¸ nªœŸ¤ °ŠÃ¡¨¸ Á°›¸¨¸œÅ„¨‡°¨
‡ªµ¤Á o¤ oœ 20% œÎ µÅžªµŠÁ¨¸Ê¥ŠÄœ£µ¡Âª—¨o°¤Á—¸¥ª„´ œ„´„µ¦š—¨°Šš¸É 1 Áž}œÁª¨µ˜nµŠÇ „´ œ‡º°š¦¸˜Á¤œ˜rš¸É 1, 2,
3 ¨³ 4 ªµŠÅªoÁž}œÁª¨µ 0, 0.5, 6 ´ÉªÃ¤ŠÂ¨³ 24 ´ÉªÃ¤Š ˜µ¤¨Îµ—´ Á„È ˜´ª°¥nµŠÄ °ŠÂ˜n¨³š¦¸˜Á¤œ˜rŞÁ˜¦¸¥¤
total RNA ˜µ¤ª·›¸ „µ¦ °ŠÁž¨ºÊ°Š (2549) ¨³ª·Á‡¦µ³®r„µ¦Â—Š°°„ °Š¥¸ œ—oª¥Áš‡œ·‡ sqRT-PCR
Ÿ¨ °Š„µ¦„¦¸ —˜n°„µ¦Â—Š°°„ °Š TDFREP241 ĜœÊε¥µŠ °Š˜o œ¥µŠ¡µ¦µ
«¹„¬µŸ¨ °Š„µ¦„¦¸—˜n°„µ¦Â—Š°°„ °Š TDFREP241 ĜœÊÎ µ¥µŠ °Š¥µŠ¡µ¦µµ¥¡´œ›»r RRIC110 š¸É¤¸
°µ¥»ž¦³¤µ– 3 že ž¨Šž¨¼„˜´ÊŠ°¥¼nš¸É˜Îµ¨¡³ªŠ °ÎµÁ£°Á¤º °Š ‹´Š®ª´—Š ¨µ Ž¹ÉŠÁž} œ¥µŠš¸ÉŤn¤¸„µ¦Ážd—„¦¸—¤µ„n°œ
×¥ªµŠÂŸœ„µ¦š—¨°ŠÂ»n ¤˜¨°— (completely randomized design, CRD) „µ¦š—¨°Š¤¸ 2 š¦¸ ˜Á¤œ˜r‡º° š¦¸˜-
Á¤œ˜r š¸É 1 „¦¸—š»„ª´ œ˜n°ÁœºÉ °Š„´œ š¦¸˜Á¤œ˜rš¸É 2 „¦¸—ª´œÁªoœ 2 ª´œ Áž}œ¦³¥³Áª¨µ 10 ª´œ š¦¸˜Á¤œ˜r¨³ 5 ŽÊε „¦¸—¥µŠ
Áª¨µ 06.00 œ. ×¥ž¨n°¥Ä®o œÊε¥µŠÅ®¨¨ŠÄœ®¨°— œµ— 15 ml š¸Én°¥¼n ĜœÊε Ȋ˜¨°—Áª¨µ ¦ª¦ª¤œÊÎ µ¥µŠÅž
Á˜¦¸¥¤ total RNA ˜µ¤ª·›¸„µ¦ °ŠÁž¨ºÊ°Š (2549) ¨³ª·Á‡¦µ³®r„µ¦Â—Š°°„ °Š¥¸ œ—oª¥Áš‡œ·‡ sqRT-PCR
„µ¦Â—Š°°„ °Š TDFREP241 Ĝ„¨oµ¥µŠ„¨»n ¤Ä®oŸ¨Ÿ¨· ˜œÊε¥µŠ¼ŠÂ¨³˜Éε
Á˜¦¸ ¥¤Âž¨Šš—¨°Šž¨¼„„¨o µ¥µŠ¡µ¦µµ¥¡´œ›»r˜nµŠÇ š¸É Ä®oŸ¨Ÿ¨·˜œÊε¥µŠÂ˜„˜nµŠ„´ œ‹Îµœªœ 8 µ¥¡´œ›»r
ž¦³„°—oª¥µ¥¡´œ›»rš¸É Ä®oŸ¨Ÿ¨·˜œÊε¥µŠ¼Š‹Î µœªœ 4 µ¥¡´œ›»r ŗo„n RRIT251 BPM24 PB235 RRIM600 ¨³
µ¥¡´œ›»rš¸É Ä®oŸ¨Ÿ¨·˜œÊε¥µŠ˜Éε 4 µ¥¡´ œ›»r ŗo„n AVROS2037 RRIT402 BPM3 ¨³ KRS138 ×¥ªµŠÂŸœ„µ¦
š—¨°ŠÂ»n¤˜¨°— (completely randomized design, CRD) ¤¸ 8 š¦¸˜Á¤œ˜r š¦¸˜Á¤œ˜r ¨³ 10 ŽÊε Á¤ºÉ°„¨oµ¥µŠ¡µ¦µ¤¸
°µ¥» 18 Á—º°œ ‹¹ ŠÁ„ȝœÊε¥µŠ‹Îµœªœ 10 ‡¦´ÊŠ„¦¸— ×¥„¦¸—ª´œÁªoœ 2 ª´œ œÎµœÊÎ µ¥µŠÅžÁ˜¦¸¥¤ total RNA ˜µ¤ª·›¸„µ¦ °Š
Áž¨ºÊ°Š (2549) ¨³ª·Á‡¦µ³®r„µ¦Â—Š°°„ °Š¥¸ œ—oª¥Áš‡œ·‡ sqRT-PCR
„µ¦ª· Á‡¦µ³®r„µ¦Â—Š°°„ °Š TDFREP241 ×¥Áš‡œ·‡ sqRT-PCR
œÎ µ total RNA ˜´ª°¥n µŠ¨³ 2 ôg Ş´ŠÁ‡¦µ³®r single strand cDNA (sscDNA) ×¥Ÿ¤„´ random primer
0.05 mM n¤š¸É°»–®£¼¤· 70 OC Áž}œÁª¨µ 5 œµš¸ ªµŠÅªoĜœÊÎ µÂ Ȋ 5 œµš¸ ‹µ„œ´Ê œÁ˜·¤ AMV reverse transcriptase 5x
reaction buffer dNTP mixture Á°œÅŽ¤r AMV reverse transcriptase ¨³ DEPC treated H2O Ä®oŗož¦·¤µ˜¦¦ª¤