full2010.pdf - page 120

82
Ÿ¨„µ¦ª·‹´¥Â¨³°£·ž¦µ¥Ÿ¨
ª·Á‡¦µ³®r¨³‡´—Á¨º°„¥¸ œš¸É¤¸„µ¦Â—Š°°„˜„˜nµŠ„´œ¦³®ªnµŠ¥µŠ¡µ¦µš¸É¤¸°µ„µ¦Áž¨º°„®oŠÂ¨³ž„˜·
‹µ„„µ¦ª· Á‡¦µ³®r„µ¦Â—Š°°„ °Š¥¸ œÄœÁž¨º°„°n°œ¥µŠ¡µ¦µš¸É ¤¸°µ„µ¦Áž¨º°„®o ŠÂ¨³ž„˜·—oª¥Áš‡œ·‡
cDNA-AFLP ץčoÁ°œÅŽ¤r˜´—‹Î µÁ¡µ³ EcoRI/PstI —oª¥Å¡¦Á¤°¦r‡´ —Á¨º°„‹Îµœªœ 24 ‡¼n ŗo™ °Š TDFs š¸É ¤¸„µ¦
—Š°°„¤µ„Äœ„¨»n¤°µ„µ¦Áž¨º °„®oŠ‹Îµœªœ 20 TDFs ¨³„¨»n¤°µ„µ¦ž„˜· ‹Îµœªœ 7 TDFs (£µ¡š¸É 2) µ¤µ¦™
re-amplified ¨³ª·Á‡¦µ³®r ¨Îµ—´ÁÅ—o‹Îµœªœ 19 TDFs ž¦³„°—oª¥ TDFs š¸É¤¸„µ¦Â—Š°°„¤µ„Äœ„¨»n ¤°µ„µ¦Áž¨º°„
®oŠ‹Îµœªœ 14 TDFs ¨³„¨»n ¤°µ„µ¦ž„˜· ‹Îµœªœ 5 TDFs Á¤ºÉ°Áž¦¸¥Áš¸¥„´¨Îµ—´ÁÄœ“µœ o °¤¼¨ (GenBank) ×¥
¥°¤¦´ homology °Š¨Îµ—´ÁÁ¤ºÉ° E value ˜ÉÎ µ„ªnµ 10-4 ¡ªn µ¨Îµ—´Á °Š TDFs š¸É¤¸„µ¦Â—Š°°„¤µ„Äœ„¨»n¤Â—Š
°µ„µ¦Áž¨º °„®oŠÁ®¤º°œ„´ ¨Îµ—´Á °Š¥¸œÄœ“µœ o°¤¼ ¨‹Î µœªœ 7 TDFs ŗo„n TDFT5-1,TDF5-2 ¨³TDF5-3
Á®¤º °œ„´¥¸œ pectin acetylesterase,TDFT13-1 Á®¤º°œ„´ ¥¸œ zinc finger protein, TDFT20-1 Á®¤º°œ„´¥¸œ severe
drought-stressed, TDFT21-1 Á®¤º°œ„´ ¥¸œ equilibrative nucleoside transporter ¨³ TDFT21-6 Á®¤º°œ„´¥¸ œ
gibberellin receptor GID1 ˜µ¤¨Î µ—´ ¨³ TDFs š¸É¤¸‡ªµ¤Á o¤¤µ„Äœ„¨»n¤°µ„µ¦ž„˜·Á®¤º°œ„´ ¨Îµ—´Á °Š¥¸œÄœ
“µœ o°¤¼¨‹Î µœªœ 2 TDFs ŗo„n TDFH2-2 Á®¤º°œ„´ ¥¸œ heat shock protein binding protein ¨³ TDFH8-1 Á®¤º°œ„´ 
¥¸œ dual specificity protein phosphatase nªœ TDFs °ºÉœÇŤnÁ®¤º °œ„´¨Îµ—´ ÁÄœ“µœ o°¤¼¨(˜µ¦µŠš¸É 1) Ž¹ÉŠ TDFs š¸ÉŤn
Á®¤º°œ„´ ¨Îµ—´ÁÄœ“µœ o°¤¼ ¨°µ‹‹³Áž}œ¥¸œš¸É˜„˜n µŠÅž‹µ„¥¸œ °Š·ÉŠ¤¸¸ª·˜°ºÉœÇ š¸É¤¸°¥¼nĜ“µœ o°¤¼¨ ®¦º °¨Îµ—´
Áš¸Éŗo °µ‹‹³Áž}œnªœš¸É Ťnž¨¦®´ (non-coding region) Ž¹ÉŠÄœ·É Š¤¸¸ª·˜š¸É˜„˜nµŠ„´œ¤´ „‹³¤¸¨Îµ—´ÁÄœ¦·Áª–œ¸Ê
‡n °œ oµŠÂ˜„˜nµŠ„´œ—oª¥Â¤oªnµ‹³Áž}œ¥¸œœ·—Á—¸¥ª„´œ„Șµ¤
˜¦ª‹°„µ¦Â—Š°°„ °Š¥¸œ—oª¥Áš‡œ·‡ RT-PCR
‡´ —Á¨º°„¨Î µ—´Á °Š TDFs š¸É¤¸ expression pattern °—‡¨o°Š„´°µ„µ¦Áž¨º°„®o Š °Š¥µŠ¡µ¦µ‹Îµœªœ 4
TDFs ŗo„n TDFH2-2, TDFT5-3, TDFT20-1 ¨³ TDFT21-6 ŞÁž}œÂÄœ„µ¦´ŠÁ‡¦µ³®r Å¡¦Á¤°¦r Á¤ºÉ°œÎµÅ¡¦Á¤°¦r
—´Š„¨nµªÅž˜¦ª‹°„µ¦Â—Š°°„ °Š¥¸ œÄœ total RNA »—Á—¸¥ª„´ š¸Éčoª·Á‡¦µ³®r cDNA-AFLP ¡ªnµ¤¸ expression
pattern °—‡¨o°Š„´  cDNA-AFLP š´ÊŠ®¤— (£µ¡š¸É 3) —Šªn µ expression pattern °Š cDNA-AFLP ¤¸‡ªµ¤œnµÁºÉ°™º°
¨³Å¤n¤¸ false positive °ŠÂ™ TDFs š¸É‡´—Á¨º°„
¥¸œš¸É¤¸„µ¦Â—Š°°„ÄœÁž¨º°„°n°œ¥µŠ¡µ¦µ˜oœž„˜· ¤µ„„ªnµ˜oœ°µ„µ¦Áž¨º°„®oŠ
TDFH2-1¨³ TDFH8-1 Á®¤º °œ„´¥¸œ heat shock protein binding protein (HSP) ¨³ dual specificity protein
phosphatase (DSP) ˜µ¤¨Î µ—´ š´ÊŠ°Š¥¸œ¤¸„µ¦Â—Š°°„ÄœÁž¨º°„°n°œ˜oœž„˜· ¤µ„„ªnµ˜oœš¸É¤¸°µ„µ¦Áž¨º°„®oŠ Ž¹ÉŠš´ÊŠ
HSP ¨³ DSP ¤¸šµšÎµ‡´˜n°„µ¦žj°Š„´œ®¦º°¨— program cell death ×¥ HSP ‹³nª¥žj°Š„´œ„µ¦¼ Á¸¥œÊ娳
žj°Š„´ œ„µ¦¼Á¸¥£µ¡›¦¦¤µ˜· (denature) °ŠÃž¦˜¸œ (Weining et al .,2002) nªœ DSP Áž}œÁ°œÅŽ¤rš¸ÉÁ¦n Šž’·„·¦·¥µ
˜´—®¤¼n¢°Á¢˜‹µ„ threonine ®¦º° tyrosine °Š mitogen-activated protein kinase (MAPK) šÎµÄ®o MAPK °¥¼nĜ¦¼ ž
inactive Ž¹ÉŠ MAPK Áž}œÁ°œÅŽ¤rš¸É¤¸šµšnŠÁ¦·¤ program cell death (Bethany et al.,2003) œ°„‹µ„œ¸Ê DSP ¥´Š
nŠÁ¦·¤„µ¦´ŠÁ‡¦µ³®r auxin ¨³ abscisic acid Ž¹ÉŠ¤¸šµšnª¥žj°Š„´œ program cell death °¸„—oª¥