Nhân giống vô tính một số loài lan hồ điệp

umo thanhnhUngkh5i nail d~mdo slf tr~mhi<$ncua tanintrong K2CrO4[8], cac cha'tn~yse theebo m~chdi xu5ngva khutch tan VaG trongmoi tru'Cfngnuoica'y.Do do,chungWiphaica'ychuyenm~usangmoi tru'ongmdieach1tu~nll~n. DS h~ncht5slftitt hqpcha'tphenoln§y, m~umangm~mdu'qcd~tc~yd vi tri ngu'qcchien. K "" ~et qua: . m~uca'ygiamtie"traphenolamdenmoitru'ong, . ti1<$m~uca'ykhongphattrieDgiamtll'33,3%con8,3%. Co thesl1giamtitt phenoltrongmoca'ykhi d~tngu'qchienla do tmhhfi'uc1.1ccuacaeproteinchuyenchdcacchc1t,rong-docohqpcha't phenolvaABA, bigiando~n.Dodo,cothesankhim~mphattriSnm~nh, m§uc3'ydu'<;1cd~tthu~nchienthls1.1ucche'cuah<;1pch3'tphenolvaABA khongdangk6mIa. -81- Phin ThaoLuJn Chung Lu?n Ail PFs. KhoaH9C Phlldngthliehoq.td9ngeuacaeeh&lieehtf10;#phenolehllaro rang,EblIngk.hiallc;tetrilnghc;tpthj caehc;tpchatphenolselien kef vc1iproteinvalieehtfhoq.td9ngeuaenzim(Salisbury,1991; Va Van V{l,1993)[50;13]. 2.VAl TROCUA CRAT DBSTTVmEN STANG TRu'<1NGvA PHAT TRItN CUA MAM NGU. Sinhtru'ongvaphattri€n la nhtmghi~ntu'<Jngra'tphuctqpchinanh hu'ongboinhtmgthayd6icuamoitru'ongvanhtmgye'uto"belltrongthlfc v~t.Cha'tBHSTTV la ye'uto'belltrongcovaitraki€m soMquailtrQngd6i vdinhfi'nghi~ntu'<Jng§y.Moi tru'ongbellngoai(xu1ycach6nh<JpKTT ngoqisinh,hqpcha'tdinhdu'ongbonchocaytr6ng,nhi~tdQ...),co anh hu'dngtrenstfphattri€n cuathtfcv~tbAngcachlac dQngtrencacphan U'ngbie'ndu'ongtrunggianbell trongcdth€ thtfcv~td€ cu6i clingtqOthanh mQtcayhoanchinh. Trqngthaingucoth€ la: . Ke'tquacuastfcom~tcaecha'tucche'belltrongm§m. . Cac v~y13baaquanhm§mkhongd&tha'mnu'dcva traod6ikhi vao m§m du'<Jc. Taiz vaZeiger(1991)[58}chothaymlieAAB tang1enmfJnhtrong phin kJn caemongu.Tuynhien,mliea9ngdeuamdmkhongphm L -82- Phin ThaoLuin Chung LuiJI An PT5. Khoa H9C dos1/hi?n di?n cuaME nhi€u hayit roa dos1/c9nbAnggiila ME vacacchatEJHSTTV khacnhdxito. vagib... KSt quado hamlu'qngchc1tDHSTTV c6 trongnhanhphatboa cho thc1y: . Hamlu'qngaux.,gib.,xito.tangtti'm~msd1dSnvi tri g~nboanod~u tien. . Hamlu'qngAAB giaml~ntti'm~msd1dSnvi tri g~nboanod~utien. Dohamlu'qngAAB cao0caevi trim~mdu'diclingnenda: Klrnh~ J! h~.- amstfnayc 01. - Uc chSstfdi chuySnxudngcuaAlA. - ABA phdihqpvdihqpchc1tphenoltrongmftuuc chSho<;ltdQngcua caeenzimc~nthie'tchostfn:Iym~m. Hayashi(1979)[trongMai Tra'nNgqc Tieng,1989J:gib. lam tangldl/ngaux.cuacacroota"baa,cdle la vi gib. cd lac dl!ng phahaycacph9nhdat6cdnhi?m VI!trongs1/tho;ilbi§n aux.. Do v~yhamlu'qngcua gib. va aux.d~ucao d caevi tri m~mtren cling. Phf!lTlEJinhTh;il (1980)[7J: gib. cdkhJ nangDch thfchquatrinh quanghl/p vahohap, RogerFrat (1993)[87]:ho/!.tdt)ngcuaaux.treEs1/keDdill fe/baa cdth€ ph9nbi?! thanh2 cdche/chinh: - Cd cha"dill h/!.fld roucat?bi€u hi?n di tmyen -83- Phin ThaoLuJn Chung LujJ1 .AuPTS. KhoaHf?C - Cd che'nglfnh;plthamd1/tr1/ctieptrendilctfnhcua vachte"baa. Phanzingchuylu nhatcuaauxin.. - Tang dt?keD dai cua te"bao. - Tang tfnhm€m deo cua vach te"bao. -Nguyensinhchattntdng1en. Aux. kich thichs1/memdeo vachIe'baaa n6'ngdt?rat thap,ldll chiulac dllngcuaaux.,cacpo1yxaccaritcuavachIe'baatra;qua biend6lgianna dos1/llinhthlmhvapha vacaclienketct?nghoa tri.N6ngdllionY tang1entrongvachte"baDnhaaux.lienket vdi caecd quailcamnhJn dilctntngcuahocman,dongval tro quail trQngtrongWellhoasinhWang(thi!qUaD)d€ Welltieldongion1-1" quamlmgIe'baa.Cae!hi!quailco th€ ho/J!hoacacbdmchudt?ngd€ din tmyenion tif Ie'baachatdenvachIe'baapilla ngoaimlmgsinh cha1.T?i daydon6'ngdt?ion }f"caoDenlampH giam.Do do,co th€ ho?thoa cac enzimclt dtfthaypha va caelien kef ngang-lien kef hydrogiila cacbos{fixen1u1ozatrongvachte'bao.Ket qua1avachIe' baam€m ra, Iranenlong1eohdn1amte"baDkeDdaira. C6 th~day1affiQttrongnhU'ngnguyennhanlambi€u hi~nki€u hlnh acaevi tri trencling1at<;10phathoafull'ea'p.CaekS"tquan~yding philh<jp vdi caenghiencU'ucuaReisingerva CSV, 1976;Tanakava Sakanishi, 1978[45.57]. -84- Iii Phin ThaoLuJn Chung Luin ADPTs. KhoaHfJC Mai Trin Ng9cTieng(1989)[9J; Salisbury(1991)[50J:xito.szl' dqngtrongm6i tru'Clngnu6icaycd lac dqng: -Fha vami~ntrfJ.ngcuacacmim. - Thlicdffysljphattri€n cuach6i - Kfch thichs1/phanchia~phanboa vagia tiingnch thu'dc uait!'baa. Phf}.mEJbJhTh;i.i(1980)[7J; TaizL. va ZeigerE (1991)[58J xito.: - Cd lac dQngrat1dndenquatrinht6nghqpprofit va axitnucleic. - CdkhaDanglam chocacchatdinhdu'angtrongcay ho~ctrongmoitru'Clngt4ptrongvgvangno lacdQngclingnhu' lam tangham1u'qngcacchata vangnly. Kef quanghiencri'ucuaPassera,1987[41J tren1liam1cho thayBAP lam tiingquanghqp do tiinghofJ.ttfnhcuacacenzim Rubis Co (Ribu1ozo-1~5biphotphatcacboxy1aza) va GPDH (G1yxeraJdehytphophatdehydrogenaza). Sankhi d~tcacmcimngutrongmoi tru'()ngnuoi ca"yin-vitrova xu 1y cacmcimnguvdih6nhqpKTT coxito.(1in-vivo,kStquanhu'san: V~quansathlnhthaigiaiphftucuamftuv~tdffdu'qcnhuQm2 mall chotha"yd vi tri meims6 1va 2 (Hlnh4), khongco phatthe13baaphu vungsinhmo.bentrongbom<;lchvftncongidih<;lnd rheincorpuschu'anot vdi ffi<;lchdftnchinhde nuoi ch6i.Songsongvdi vit%cphantichham1u'qng -85- Pha'n Thao Lu#.nChung Lu;!.l:1All PTS. KhoaH9C cha'tDHSTTV nQisinhchotha'ydnhifngvi trinAyhamluqngAAB caoda klrnhamstfnffych5i,ucchSslfdi chuySnxu6ngcuaAlA. Do d6,khixuly xito. voi mt;lcdich tc}'ocho corpusphancilia theomQihuang,khi dC}.tkich thu'oct6i da,baa ao ngoaitachdATIva ch6psinh tru'ongphattriSn thanh ch5iv~nkhongdC}.tdu'qckStqua. dnhanhphatboachu'anovadnhanhphatboadanohoanloancho tha'yd phatboadano hoanloanthl du'C1ngkinhcQngphatboaIon hdn, chiSudftycualopnhumo,s6lu<;1ngcuab6gSva b6 libe nhiSuhdn.Stf hlnhthanhnhi~ucacb6mC}.chg6pphAnv~nchuySncaccha'tdinhdu'ong vac:k cha'td6nghoadSnuoimAmsannAy.Hdnnua,ciiysankhi tr6boa tmchungtabiStdu'qcmallsAc,cha'tlu'qngboacuacaymymQtcachchmh xac , day1avit%clamfa'tcftnthie'tkhi dungcaymy dSnhangi6ng. v ~biSuhit%nhlnhthaibellngoaichungc63 dC}.ng: - NIAnlkhongphMtriSn(vi tri mftm1va 2). - Choidinhdu'ong(vi trimAm3va4). - Ch5isinhsan(phatboathll'ca'p),lanhifngmftmn~mdbelltren. KSt quanAyphuhqpvoi nh~nxetcuaTanakava Sakanishi(1978) [57],BouriquetvaBroly(1982)[17]khi thlfchi<$ntrongmoitru'Cfngnuoi ca'yin-vitro.KSt quatren1ado: . Slj phanbbcaccha'tBHSTTV dtU'ngvi trimAmngutr~ngnhanhphM boamachungtoidaphantichdtren. . Ben cC}.nhdo.anhhu'angcuatinhtrQidinh(apicaldominance)kiSmsoM codi~uchinnsqsinhtru'dngvaphattriSncuacacvi trim~mtrennhanh -86- PhffnThaoLu~ Chung Luin Ail PTs. Khoa H9C phMboa(dotmhdi chuy~nhifucrfccuaaux.).Auxin du'Qcsinhfa 111'n~ ngQnc6th~di chuy~nxu5ngcacvi tri mgmbell du'disailkhi C:ltbo n~ ngQn,vi tnmgmggnngQntillhamIu'qngaux.caohdnd nhiTngvi tn mgmke"titpdotinhdichuy~nhifucrfccuaaux.,dod6 tmhkeodaitt baac6u'utht hdnliend:It~ophMboathuca'p. NjshimuraG.~1972;TanakaM~ 1978[39~57J:cacmim nguchiu iinhhu'cJngriftnhieubcJjnhitftdQva vi trimim trenphathoa. Trongnuoiciy m-vjtrokhi nhi<$tdQthip khoang20oeti 1<$ch6isinh san tangkhoang86%,1rongkhi d nhi<$tdQcaokhoang28°Cchi c6 8,3% vavi tri xuit hit%ncnaphMboathuca'pla nhfi'ngvi tri bell trenggnboano dgutien. Phu'dngphap giam canh trongnuoi ca'ym-vjtro 111'ch6i sinh san khongmangl~ihit%uquanhangi6ngcaovi khi C:ltdo~ngiaml~ichungchi cho ra ch6i dinh du'ong.Nhu'v~yht%s6 nhan gi5ng cling khong cao, Bouriquetva CSV, 1982[17]clinggill nh~ntu'dngtIt nhu'tren.Trndc nhifngkh6khang~pphai,d~hit%uquanhangi5ngtrend09nphMboadu'qc cao,chungtoi tie"nhanhnuoica'ydinhngQncnanhiTngmgmngungyvdi m~cdicht90nhiSutiSncn(protocorms),ph~cv~chonhangi5ng.DiSungy d:I du'qcthtjchit%nbdi TSE va CSV, 1971[62] tren gi6ngBia Ian (Cymbidium).Ngoai ra vit%cnhangi6ngtU'dinhngQncon t90du'qcgi6ng s(;lchbt%nh,khongbi nhi~mvirus(AugeR., Beauchesne,1982)[69]. Ke"tquanuoi cffydinhngQnmgmngn cna phM boa (Bang 3), cho thffychi c6 a vi tn mamso'4 va sO'5, ti It%phM tri~nthanhnhffngkh6i u -87- j . ... ::I.III i PhanThaoLutln Chung Luj11A1zPTS. KhoaH9C nho baoquanhdinhmftmgiatangtrongmoitru'dngnuoic~yMS cothem 2,4D.Cha"tnftythu'C1ngdu'<;1eSITd1JngnhiSua ddntITdit%pnhu'Iua,chu5i, caeIo9icolamthucangiasue(LinaceroR.,1986;Da-YuanWang,1982) [32;22].Khi m~uea"yIa tiSnen5-6thang,moitru'CJnguoica"ycochua BAP va2,4D tmu'uthe'ngQncnam~uea"ysebi ueehe'vaehofamQtdam te'baophattri~ntheoehiSungang. Ngoaianhhu'angenacaeehltDHSTTV nhu'd?ikStrentilltheo: Uratava Iwanaga,1965;Scully, 1966;Tanakava Sakanishi, 1978[63,53,57J th1sl/themnu'dcdiravaomoi Wangnuoicay in-vitrovdiDongdQthichhc;lpco th€ thucdayho~ttfnhczlachol bell c(fngnha'sl/hinh thanhcaycon. Caethanhphftntfongnu'dedU'acoho9ttinhneh thiehslfphanehia te'baogi5ngnhu'ho9ttmheuaBAP, dodocothSthaythe'du'<;1eBAP. Theosl/ph/in tfchthanhphffndinhda'angtrongna'dcdiraczlaT6 chlfc Y te"The"gidi (WHO), V(f viln V~va Csv, 1993[66.13], trongnu'dcdiraco kha giau cachc;lpchatnitd d~ngkhd'nhu'cac axitamiD,cachocmonsinhWifng nha'xito.. Dodo,khichothemvaomoitru'dngnuoiely seb6sungngu6ndinh du'ongIonvachltBHSTTV c<1nthie'tehoslfphattriSncnam~uv~tca"y. -88- Phdn ThaoLuqn Chung Lu~ Ail PTs. KhoaH9C 3. ~ANG TAOKHOI MOvA TAJ SINUKHOI MO. DayIadi~mmdicuaIu~nan,chu'acangtrlnhnaocangbo'. Khi nehthichmftuca'ybAngnhiffigve'tdamnho0ph~ng5cthancay d~t<;LOthanhve'tthu'dng,daphatsinhnhiffig kho'ite'baava t5ehU'c0 trong mai tntongnuaiea'yc6neh thieht6thiehh<;fp. Halperin(1969);Nozeran,DureuxvaRossignol(1982)[84]: - Caete/baDnaysechius1/phanphanh6atntdclin phanchiadall lien. 31/phanphanh6a d6ngvai fro quan1r9ng,tlY1 te/baDtntang thanhcua1 cdquancdth€ quayve'lrf!J1gthaiaunien. - 31/Ireboanayc6 th€ tMsinhranhilngcdquanho~ctf!Ophoi dos1/ cdm~tcuachatEJHSTTV Irongmoi tntCingnuoi cajl. Bili TrangVit;t(1994)[88]s1/sinhphoi tii'te"baathe"ht; c6 th€ qua2 giai dof!J1: . Giai dof!n1: Giai dof!ncamling s1/tf!Omo St;Ocdkha nangsinh phoi(calsembryogeneses). £)613nhfi'ngnh6mte'baakhangphanboa do sQ'sinhsantt baacua matrongnuaica'yin-vitrovdi sIthit%ndit%ncuaaux.riengIe haykSth<;fp vdixito. PhftuthU'ccAtngangph~nthan0 cu5i g6echo tha'ysIt t<=;1Okh5i ma xua"tphattti'vungchU'acacb6m<=;1ch.C5 ph~ngoai,sQ'phanciliat6baaxay fa IQnxQnkhangcot5chll'e. -89- Phln ThaoLuJn Chung Lujo .AnPTS. Khoa H(,JC II Giai doflJl2: Giai dofl.nbi§uhitlnquatrjnhsinhph6i . NhlingkhO'imosailthdigiannuoicf{ytrongmoitru'dngc6aux.dac6 stfphanh6ad€ t90chdi,dangthditrongmoitntdngSITdt}ngc6xito.da nch thichstfbi6nd5iti~n19Pthanhc:k di~p19Pc6h09tdQngchU'enang: - C6nhi~u16khidu'qehlnhthanh. - C6 nhi~uh9ttinhbQtd9ngiron,nho,c6 Ie caeh9ttinhbQtnfiydang trongquatrlnhphanbuyd€ sudt}ngchocaeh09tdQngeuat6baa. K6t qua:dkhO'imotai sinh,hamlu'qngDLT giatang,CBQH gia tang,c6 stfgiatangt5nghqpcaeprotein d€ hlnhthanhmdi caecdquail. Sau90ngaynuoici1ytrongmoitru'dngc6chU'aux.,ti lt$caytaisinh caodmoitntonge6ehU'a3mg/lANA, c6stfhlnhthanhcdquailla var8d€ 190thanhcaycon. 4. VAI TRO CUA CRAT KHOANG TRONG MOl TRU(JNG DINH DUONG. Caemoitru'dngdinhdu'ongdu'qesudt}ngtrongnuoiei1yin-vitrodS t90cdquail,khO'imo,r8vatrongdungdichdinhdu'ongtu'diehocaytrdng lamoitru'dngMurashigevaSkoog,1962. K6t qua:mfiuv~tei1yphattriSnto'ttrongnuoici1yin-vitrova cae chdicon,caycontangtru'dngvaphattriSn16ttrongnuoiin-vivo.AugeR. vaCSV, 1982[69]clinggillnh~nhu'tn~n,c61eVI: . Ngu6nclingcHpd9mtrongmoitntdngcao. -90- Phin ThaoLuJn Chung Lu$nAuPTS. KhoaH9C . C6 slf canbAnggifi'aanionva cationtrongmoi tru'Clngdo d6 thlchh<;$p voi mfiuv~tca'ytrongvi~cduytrl apsua'th5m tha'ucuamo. C5 hq.td~leu m§mdu'<;$ctmba'tleYhq.tnaoclingdSuphiii tdHquacac giai doq.n,b~td§u 13hq.tthu nu'ocdS boa tan cac cha'td phoi nhli, hoq.th6a" enzimtq.odiSuki~nchocacphanlingthuygiaidi~nramq.nhmeclingca'p nguyenli~uchohoq.tdQngbie'ndu'ongnha'tla h6ha'pd~t5ngh<;$pcha'tmoi nhu'protit,lipit,gluxitc§nthie'tchoslftangtru'dngcuacaym§m(JohnA. Bryant,1985;Taiz L. vaCSV, 1991)[25,58].Khongnhu'0 hq.t,cacvi tri m§mngii 0 nhanhphatboaphattri~nsankhi xU'ly cha'tBHSTTV ph~ thuQchoanloanvaolu'<;$ngcha'tdinhdu'ongva nu'occlingca'ptrlfctie'pcho m§mva thongquah~th5ngr~dSchuy~nleu Quaim§m.Do d6,songsong voi vi~cxU'ly cha'tkkh thlchm§m,chungtoi phuncacdung.dichdinh du'ongkhacnhaunhAmtlmmQth6nh<;$pdinhdu'ongkhoangtruchh<;$pnha't dScaccayconsinhtru'dngvaphattriSnt6tva duytrl t6tdClis6ngcuacay my,vllan H6Bi~pla mQtloq.icays6ngbi~usinhcholienvi~cclingc.1p dinhleYcaccha'tdinhdu'ongkhoangla r.1tc§n thie't. Sankhi tu'oicacdungdichdinhdu'ong,chungtoi nh~ntha'ycacke't quado cu'ongdQquangh<;$pva cu'ongdQhoh.1psan1tu§nva 2 tu§nd cac moi tru'Clngdinh du'ongdSutangtll'vi tri m§m so'1 de'nm§m g§n boa nd d§utieD(Bang14,Bang15)vatangcaohdnsovoi d6ichling.Tuynhien, d moitru'ClngMP2 ke'tquadoCBQH vaCBHH caohdnnhiSusan2 tu§n tu'oidungdichdinhdu'ong(BanI!16),VI trongmoi tru'ongn§y co ham lu'<;$ngdq.mcaovacothemkhoangvi lu'Qngnhu'MgSO4vaZnSO4. -91- Phifl ThaoLuJn Chung Luin An PTs. KhoaHfJe Natre (1970)[82J; David T. (1985)[20J: ham1u'{fn~d;pncaDtrong caccd qUaDd6'ngh6alam tiingcu'Clngdi?quangh{fpvangu'{fc1?is1/ hinh thanhvahOlJ!di?ngcua bi?may quangh{fp1?i lien quanchijt chi!vdimzfcdi?cungcapd;pn. Zn c6lacdlJnglamtangkhaDanggill nildc,di?ng4mnildccuamo dolamtiingquatrlnhsinht6ngh{fpcaecaDphantll'ila nildenhil protein,axitnucleic. Brian WilJjamsvaJackKrammer(1983)[91J:S1/thieuMg dijcbit;t d61vcJi1ananhhlidng den s1/tf!.othanh dit;pllJc to"d la, do d6 cay d6 hinhi6mflambt;nh.NghiencrfucuaBraunvaSteinboc,1953trong [13J cho thayMg 1a tac nhanhof!.thoa hang1of!.tenzlinnhli photphataza,eno1aza,kinaza xdctacchoquatrlnhphangiai cac nguyenlit;uhftucdtrongquatrlnhhohap. Nhu'v~ycaecha'tkhoangcoanhhu'angtntctitp dtn quatrlnhtrao d6icha'trongcay. Sq'thamnh~pva v~nchuyencuacha'tkhoangclingphI}thuQcvao CBQHvaCBHH (Ph~mBmhThai(1980)[7];Vli vanVI}(1993)[13]. Trongmoitru'CJngMP2, ham1u'qngd~mt6ngs5trencaymy sau2 tugntu'didungdich dinh du'ongcaohdn h~nso vdi cac moi tru'CJngkhac. Phantichphu'dngsaichotha'ysq'khacbi<%t1acoynghlaarillIC 99%. Ktt qua:slf tangtru'dngva phattrieDcuach6i cont6t,xua'thi<%n1a va r~nhanhhdn. -92- Phin ThaoLuQnChung LU?l1 An PTs. Khoa H9C Mai TrinNg9cTieng(1989)[9];Nguy6nVanUy& .{1989)[12];Vl1 vanVqvaCSV (1993)[13]:Sifhinhthanhr6 cuacacc§yconnhanh 18dosifd6ngg6pcuaZn vi : - Zn c6Hen qUaDdenham1u'ljngtriptophan-ammoaxit,ti€n thJn cua quatrinhsinht6nghljp AlA 1ft d6 thucdly nhanhsif l;;tor6 cuac§y con. - Zn c6 vai tro trongvi?c thucdly sif vQnchuytfncacsanphim quanghljp Ill'1aden cac cd quaD. VI v~y,khi phantichhamlu'Qngaux.nQisinha cay contill moi tru'dngMP2 choktt quacaohdn. Phftuthucc~tngangndit~osdkhairSg6m: . Nhi~udamt€ baadangphan cilia m~nhtll'lop cac tt baabo m<;J.ch hu'angfa ngoaiyO,d6nepcact€ baava eplen cacbo sQicu'dngmo. . SanmQthdigianchungn6ili~nvoibomQccuarSsaunfiy. . C5 vungchoprSlanhifngtt baamomSmcochuatinhbQt,choprSnfiy duQcbaophilbAngmQtlop nhfiy,lop nfiyco th~tho di thanhIDQtlopvo cung.TheoWeber(1953)[trongKatherinEsau,1980][2],dayla rSkhi sinh. Nguy6nNg9c TJn, Nguy6nEJinhHuyen(1981)[1]: Sif tangmdc aux.trangcdquannaod6sethucdly dongchatddnghoaquanghljp vanhitngmu61dinh du'iJngill VaGtrangcd qUaDd6. Ktt qua cacch6iconphattri~ncand6iva khoe,bQrS t6tco th~ tachtr6ngvaoch~urieng. -93- J ._.