C CH HP TH ION CA NHNG T BO RING L V R

Nhng th nghim khng nh v tnh ton s tp trung ion (mM) da trn s tch in.

in th trong r ca u H Lan v yn mch

Nitrate R u H Lan( -110 mV) ION KALI NATRI Thc nghim 75 8 28 0.027 R yn mch (-84mV) Tnh ton 74 74 Thc nghim 66 3 Tnh ton 27 27 56 0.076

CANXI
CLO

2
7

10800
0.014

3
3

1400
0.038

Kt cu ca dung dch ngoi: 1Mm KCl1 , 1mM Ca(N03)2, and 1mM NaH2P04. Cn c vo Higinbotham er fl/. (1967). S tp trung anion trong cc khng bo da trn s tnh ton theo s cn bng ha hc. V s tp trung anion trong cc khng bo thc cht c tm ra t cc thc nghim.

Trong v d ny, Cation duy nht m s c yu cu vn chuyn ch ng cho s hp th ca n l K trong r yn mch. bn ngoi K tp trung t, vn chuyn ch ng c i hi thng xuyn. (Cheeseman and Hanson, 1979). Ni c th cho Na and Ca, s tp trung cn bng mi trng lng ca t bo (i.e. tnh c) s cao hn nhiu, iu ny c tm ra trong thc nghim khi trng thi n nh (bng 2.8). S gii thch hp l cho s tri ngc ny l tnh bn vng ca mng t ba b gii hn bi nhng ion trong hay nhng ion c bm (vn chuyn) tr li bn ngoi dung dch. Na c bm tun ra mng t bo ca t bo r c thit lp cho nhng loi cy c bit khc nhau. (Jacoby and Rudich, 1985; Schubert and Lauchh, 1988). S y ra (Ca bm tun ra) mng t bo cng tn ti trong t bo r (Gibe and Sommarin, 1991).

Vai tr quan trng ph bin ca s bm tun ra Ca c mng t bo v tonoplast cho s hot ng ca t bo c tho lun trong phn 8.6. V s tp trung Ca trong dung dch t thng cao hn 1Mm, s bm tun ra Ca ngn Ca vn chuyn cng vi gradient in ha (bng 2.8) s c mt nhu cu nng lng ng k. C kh nng, v th, thm vo nhn t ha l nh l ln hay cht mang ca Ca hn ch s thm qua mnh m cng vi gradient in th in ha bn kia lp mng. Trong nhng nm gn y, s tin b vt bc trong s hiu bit c v c ch ch o thng tin ca in th pha bn kia ca mng v s quan trng ca in th cho t bo pht trin v thc hin chc nng. S tin b l hp l bi cc phng php k thut mi cho bit kch thc ca mng in th v nhng ion i qua c thot ra t l hng ca mng hoc t nhng lt ct ca mng ( ming v k thut, Hedrich and Schroeder, 1989).

Phn ch yu ca s vn chuyn ion trong mng c trnh by trong hnh 2.8. Bm H+, tng hp ATP (sc mnh chuyn ng proton; Poole 1978), vn chuyn H+ xuyn qua mng t mt trong n mt ngoi, v d nh trong mng t bo t cytoplasm (t bo cht) n apoplasm ( ), to thnh mt gradien v pH v in th in ha. S vn chuyn ca cc cation v anion cng vi gradient c gin tip chn bi nhng ion cht mang hoc knh.

Internal signal : tn hiu bn trong External signal: tn hiu bn ngoi Hnh 2.8 C ch chnh ca vn chuyn ion trong mng sinh cht. (A) bm H+ ATPase; (B) knh ion, (C) cht mang, (D) kt cp protein cho tn hiu ca s thu nhn v s truyn tnh trng. (Thay i t Hedrich etal, nm 1986, vi s cho php t Xu hng trong khoa hc Sinh ha)

M hnh ny a ra mt chc nng quan trng khc ca mng t bo, c th l ca mt th th ca tn hiu bn trong v bn ngoi (v d nh pH, nng ion) v s chuyn i ca nhng tn hiu ny trong cc qu trnh vn chuyn ca mng.

 cng c s thng nht trong cc bm proton ca thc vt nm trong c hai mng plasma v tonoplast, v chc nng chnh ca chng l iu chnh pH trong t bo cht (Kurdjian v Guern, 1989). Nhng my bm vn chuyn H+ t t bo cht xuyn qua cc mng t bo vo apoplasm hoc thng qua cc tonoplast vo khng bo dn n s khc bit pH in hnh gia cc ngn. Theo quy nh, pH ca t bo cht (cytosol) l 7,3-7,6, khng bo 4,5- 5.9 v apoplasm khong 5,5 (Kurdjian v Guern, 1989). Theo , trong t bo thc vt s i ra cc proton t t bo cht qua mng t bo v tonoplast l qu trnh cung cp nng lng s cp v cung cp ng lc cho s vn chuyn nng lng th cp ca cc cation v anion dc theo gradient in ha.

Cc chc nng v v tr ca bm proton v vn ti ca cc cation v anion qua mng bo tng v tonoplast c tm tt trong mt m hnh trong hnh. 2.9. S vn chuyn mng t bo ca cc amino acids v ng theo nguyn tc tng t, tc l, n c iu khin bi bm proton ca lp mng (mc 5.4.1).

Trong m hnh th hin trong hnh. 2.9 mng plasma b rng buc H+-ATPase ng vai tr nh mt cha kha quan trng trong c s iu chnh ca pH t bo cht v ng lc cho hp th cc cation v anion. Do n c coi l enzyme ch o(Serrano, 1990). S tin ng k trong s hiu bit lm cch no iu chnh cc hot ng ca enzyme, cng mc gen (Palmgren, 1991). Nhng hot ng ca mng sinh cht H+-ATPase l c bit cao lng ht ni tiu th 25-50% ATP ca cc t bo (Felle, 1982). Theo , mc carbohydrate thp trong r khng ch lm gim c ch hp th ion ca nhng t bo ring l v r

Fig. 2.9 M hnh chc nng v v tr ca bm proton electrogenic (H+-ATPase, PPiase), bm mng oxi ha kh (NAD (P) oxidase), cc knh ion, v vn chuyn ca cc cation v anion qua mng plasma v tonoplast, y proton vo mi trng bn ngoi m cn lm gim pH cytosolic(Schubert v Mengel, 1986). Cc mng plasma H+ATPase c kch thch bi cation n tr theo th t K+> Na+ v n tng i khng nhy cm vi anion (O'Neill v Spanswick, 1984). Mt v d v s kch thch bi K ^ c a ra trong Hnh 2.6. Calcium ni chung v cytosolic Ca+ t do tp trung ring l ng vai tr quan trng trong s kch thch ca enzyme ny. S gia tng cytosolic Ca t do lm tng hot ng ca plasma mng enzyme, kch thch ti a xy ra ngay c gi tr thp hn 7 juu Ca t do (Schaller v Sussman, 1988).

 kch hot cn c tc ng ca Ca t do trn mt protein kinase lin quan vi cc b mt bn trong ca mng bo tng (mc 8.6.7). Cation c vn chuyn xung di, dc theo gradient in ha (khong -120 n -180 mV) trn mng t bo vo t bo cht trong uniport, cu trc c th (cht mang carries, permeases) trong mng t bo (Hnh 2.9). i vi kali (K ), tuy nhin, nng thp bn ngoi (<1 mu) th i lc cao h thng hp thu hot ng chng li s khc bit v gradient in ha (v d 10 UMK bn ngoi; -8 0 mM K hot ng trong t bo cht) Nh vy, K c yu cu vn chuyn nh cht mang trung gian hoc l vn chuyn proton antiport , nhiu kh nng l mt K+ 01:01 -proton symport, hoc cotransport (Kochian et al, 1989.;Maathuis v Sanders, 1993). i vi anion vn chuyn qua mng bo tng cotransport proton-anion (hoc symport) bng cch s dng s chnh lch gradient in ha lm ng lc.

C nhiu quan im khc nhau v c ch vn chuyn qua mng, tuy nhin nhng quan im ny cha r rng Ngoi ra cn c quan im khc v c ch vn chuyn anion qua mng plasma. Theo Liu (1979), s hp thu phosphate ca r ng c qua trung gian OH / phosphate countertransport trong vn chuyn ca OH t ni c in th cao xung thp (pH cao v in tch m mnh m)

Tuy nhin, bng chng thc nghim th trao i, vn chuyn OH hoc HCO3, qua mng l ng lc cho vn chuyn anion l yu, cht c ch c th ca OH v HCO3 khng c sn (Kurdjian v Guern, 1989). Haimy bm c thnh lp, H+-ATPase v pyrophosphatase v c, Ppjase (Hnh 2.9). C hai my bm proton phosphohydrolases s dng mt trong hai ATP hoc pyrophosphate v c l ngun nng lng (mc 8.4). Magi l cn thit cho c hai my bm, cho thy rng Mg ATP v Mg PPI l cht nn. Pyrophosphate v c c to ra trong con ng sinh tng hp vi mt s lng ln (Rea v Sanders, 1987) nh tinh bt tng hp (mc 8.4) hoc sulfate hot ha (mc 8.3.2). Nng PPI trong bo tng c gi nh l trong phm vi ca 50-390 l i bm khin proton (Chanson et al, 1985).

S tp trung ca PPiase proton bm ti tonoplast c v l thp hn ca ATPase, v c hai my bm ny khc nhau v b nh hng bi cc cation v anion v c (Bng 2.9). Ngoi tr cc thit yu ca Mg++ cho c hai my bm, Mg-ATPase c kch thch bi clorua v c ch (hoc khng b nh hng) ca kali nitrat trong khi ngc li l b nh hng i vi cc Mg-PPjase. H + bm Mg-PPjase bt buc phi ph thuc vo s hin din ca K,kch hot K xy ra giai on t bo cht (Davies v cng s, 1991b).

Cc my bm proton ti tonoplast c yu cu gi cytoso Hc pH cao v ng thi cung cp ng lc cho vn chuyn cation vo khng bo l countertransport (hoc antiport, hnh 2). i vi anion cc l cha r rng. Giao thng vn ti cc Anion t t bo cht vo may khng bo theo gradient electropotential t tiu cc trong khng bo so vi cc t bo cht (Hnh 2.9). Trong khi , s tn ti ca hai my bm proton ti tonoplast (ATPase; PPiase) v H ^-ATPase ca mng t bo cng c thnh lp, n vn l mt vn tranh ci liu mt h thng th hai ca s chuyn proton qua plasma mng t bo cht apoplasm c tham gia vo s hnh thnh v duy tr electropotential mng v pH (Hnh 2.9).

H thng c lin kt vi mt chui oxy ho kh vi NAD (P) H l ni cung cp in t. C l tt bng chng cho s tham gia ca h thng ny trong tng trng tng cng auxin do (Morre et al, 1988.), hot ng khng khun (Dahse et al, 1989), hoc gim ca Fe (III) b mt ca mng plasma gc (Bienfait v Liittge, 1988). Tuy nhin, do , n nay ch yu l ferricyanide hoc cc hp cht nhn to c th c s dng nh in t cht nhn t c mt nng lc vn chuyn qua mng ng k proton (Luster v Buckhout, 1988). Vai tr ca h thng oxi ha kh proton qua mng gradients v vn chuyn ion vn cn c vn cho n khi mt ngi chp nhn electron sinh l c th c tm thy (Serrano, 1989).

Theo quan im ca tnh hiu qu ca ascorbate (ascorbic gc t do) nh in t chp nhn cho bm mng oxi ha kh (Gonzales-Reyes v cng s al, 1992.) v nng ascorbate tng i cao trong apoplasm l (PoUe et al, 1990) vai tr ca h thng ny trong l vn chuyn mng bo tng ca cc ion v cc cht ha tan khc khng th c loi tr (Chng 3).

Gn y hn, s tn ti ca cc knh ion trong mng t bo thc vt c thnh lp v, do , cc knh c bao gm trong cc m hnh hin ti ca mng vn chuyn ca cc ion (Figs. 2.8 v 2.9). Cc knh ion l duy nht trong s cc protein vn chuyn trong ca h kh nng iu chnh hoc ch thng lng ion 'ca' mi trng vt l-ha hc ca cc knh protein (Blatt v Thiel, 1993). Cc knh cho php vn chuyn (uniport) cc ion qua mng. Knh m xc tc cho cc thm thu ion mi giy l t nht ba (Tester, 1990) hoc thm ch nm ln (Bentrup, 1989) . Tuy nhin, knh ion c ng hu ht thi gian, v s lng mi t bo c v l kh nh.

V d, trong mng bo tng ca cc t bo l khong 200 K+ trong knh ca mi t bo l gi nh (Kourie v Goldsmith, 1992). Cho n nay, cc knh c th cho K, Ca , H v Cl c xc nh, v cho knh NO c cng nhn ti tonoplast (Tyerman, 1992). C nhiu gi nh v chc nng ca cc knh ion trong t bo thc vt