GII THIEU VE TAI LIEU Ti liu bn ang xem c download t website WWW.AGRIVIET.COM WWW.MAUTHOIGIAN.ORG Agriviet.comlwebsitechuynvnngnghipnilinktmithnhvin hotngtronglnhvcnngnghip,chngtithngxuyntng hpti liuv ttc cclnhvcclinquannnngnghipchiascngttcmingi.Nutiliu bncnkhngtmthy trongwebsitexinvuilng giyucuvban bin tp website chng ti c gng b sung trong thi gian sm nht. Chngtixinchn thnhcm nccbn thnhvin gi ti liu v cho chng ti. Thaylicmnntcgibngcchchiaslinhngtiliumbnangccng mingi.Bncthtrctipgi tiliuca bnlnwebsitehocgivchochngti theo a ch email Webmaster@Agriviet.Com Lu:Mitiliu,hnhnhbndownloadtwebsiteuthucbnquyncatcgi, dochngtikhngchutrchnhimvbtkkhacnhnoclinquannni dungcatptiliuny.XinvuilngghirngungcAgriviet.Comnubnpht hnh li thng tin t website trnh nhng rc ri v sau.Mtstiliudothnh vingivchochng ti khng ghi r ngun gc tc gi, mtstiliucthc ni dungkhngchnhxcso vi bn ti liu gc, v vy nu bn l tc gi ca tp tiliu nyhylinhngayvichngtinucmttrong cc yu cu sau : Xa b tt c ti liu ca bn ti website Agriviet.com. Thm thng tin v tc gi vo ti liu Cp nht mi ni dung ti liu www.agriviet.com +~ .~+ :: ++- \ ~. PHAN I: GII THIEU Trong nhng nam gan ay vi chnh sach m rong au t ve moi mat cua Nha nc, i song vat chat va tinh than cua ngi dan ngay cang c cai thien va nang cao. Song song vi nhu cau ve vat chat th nhu cau tinh than ac biet khong the thieu c. T lau con ngi a thch trong hoa, cay canh va e trang tr cho ep, va e giai tr tinh than. Moi ngi thch trong mot loai hoa khac nhau, viec la chon loai hoa nao thng tuy thuoc vao ieu kien kh hau cua vung o, ve ep cua hoa, cung nh la hoa o co de trong va cham soc hay khong. Hoa lan c nhieu ngi a thch, bi le hoa lan co cau truc kieu k va phc tap, nhat la bo phan moi co nhng net cham tro rat tinh vi, lai rat phu hp vi ieu kien kh hau cua Viet Nam, de cham soc va moi ngi co the chon loai hoa lan mnh thch tuy theo tui tien cua mnh ma van thoa man c thu vui tao nha.Phong lan nc ta rat phong phu va a dang, co nhieu giong khac nhau nh: Cattleya,Phalaenopsis,Oncidium,Mokara,Vanda,Dendrobiumchungeuchohoa rat ep va mang nhieu mau sac khac nhau. No co the dung e trang tr, trng bay, lam ep, dung trong cac buoi le hay ngi ta co the ban hoa cat canh-kinh doanh. Trong so o co le Dendrobium la giong ac sac nhat t mau sac, dang hoa cho en giong loai. Mac khac, Dendrobium cung rat de trong, rat sieng hoa va lau tan. Do o no rat c achuongvactrongphobiennhatnctahiennaynhamphucvuchonhucau cuoc song. Cacnhatronglankhongngngtmkiemcacgionglanmiethoamans hieu k cua mnh hoac co the em ban. Co hai cach e co c giong lan mi. Mot la, sutapnhnggionglanhoangdaitrongrngemvethuanhoa,taocacieukien nhan tao giong t nhien e cay lan co the ra hoa. Phng phap nay gap nhieu rui ro do ieu kien moi trng khong thuan li cho cay lan phat trien. Hai la, tao ra nhng giong lan lai mi, cay lan lai se mang nhng ac tnh tot vt troi cua ca bo me, co the thoa mancnhucaucuangithngthclan.Tuynhienkhihaicaylanlaivinhau at ket qua va tao trai can phai ket hp vi phng phap gieo hat trong ong nghiem e hat lan co the nay mam de dang. Co nh vay mi co the kiem tra ket qua cua viec lai hai cay lan. Xuatphattnhucauthctien,trongphamvietainaychungtoitienhanh nghien cu: Nhan giong lan Dendrobium bang phng phap gieo hat in-vitro. . .y-+ .. :+ ~+ y-+ ~+ 1 +~ .~+ :: ++- + .~+ :~ -. . .y-+ .. :+ ~+ y-+~+ 2 PHAN II: TONG QUAN TAI LIEU I.Gii thieu ho lan: 1.1ac iem chung: Cac ho lan c anh gia la mot trong nhng loai hoa cao cap trong vng quoc thao moc, bao gom hn 25.000 ngan loai khac nhau, cung vi nhng loai mi c kham pha va mo ta theo tng nam. Do bi chung c phan bo vung rong ln, trai daitngxchaochoenBaccc,tongbangchoencacvungnuibang tuyet,cacloailanratkhacbietnhau:Lanat(phattrienmoctrongatkhang nc); thc vat bieu sinh hoac thc vat phu sinh (phat trien pha tren mat at hoac songbamtrencacloaithaomockhac,thuhutchatdinhdngvanctmoi trngxungquanh);thcvatphattrientrenmatahoacngaycadimatat (phat trien di be mat cua moi trng cay trong). NhngnhasanglapnganhLanhocangkelatrietgiangiHyLap Theophrastus(372-287trcCongnguyen)vasaunaylanhathcvathocngi Thuy ien Linnaeus (1707-1778). Chnh Theophrastus la ngi au tien s dung t Hy Lap Orchis e ch nhom Lan [4]. 1.2ac iem hnh thai: 1.2.1C quan dinh dng:Gia hanh (than gia): ch xuat hien tren cac loai lan a than. Gia hanh la bo phan rat can thiet cho s sinh trng va phat trien cua lan. Gia hanh tuylathannhnglaichadiepluc,aylabophandtrnhieuchat dinhdngratcanthietchosphattriencuagiahanhmi.Giahanh cung la c quan d tr nc[11]. Than:Thanvaygiaconhieuhnhdangkhacnhautuytheogionglan. Tren than co ot, tren moi ot moc mot nhanh la hoac la bao. Than la c quandtrncvachatdinhdng,mamhoavamamlaeumoct phangoccuabophanthanre[12].Chcocacloainthanvamotso loai cua giong Dendrobiumva Epidendrum va co gia hanh, va co than. Cac loai lan co than thng khong co c quan d tr nc va chat dinh dng[11]. La: la c quan dinh dng cua hoa lan, la xng che tao chat dinh dng bang quang hp. Phien la thng co hnh li kiem dai, so lng va hnh dang la khac nhau tuy chung loai lan khac nhau [12]. La co the moc oi xng qua gan chnh hay khong, la sat nhau goc hay xep cach co be up lennhau,chiaoteuan,cokhithoaihoathanhvayhayphnhlen, mong nc, hnh dang rat khac nhau [11]. Canhanh(than-re): ch gap lan a than. Can hanh la ni cau tao cac cquandinhdngmi,trencanhanhconhieumatsong,chethoac +~ .~+ :: ++- + .~+ :~ -. . .y-+ .. :+ ~+ y-+~+ 3 hunien.Matlanihnhthanhcu ngmangratnhieureenuoisong cay lan [11]. Re: lan a than, re thng c hnh thanh t can hanh. cac loai n than th re moc thang t than va thng xen ke vi la. Re tren khong cua cacloailanphusinhcomottrucchnhbaoquanhbimokhongchat, giong bot bien bao quanh goi la mac. Mac co the hap thu hi nc cua khong kh, cung nh tch tr nc ma va sng ong [11]. 1.2.2C quan sinh san: Hoa:taphpthanhcum hoa chum hay bong. Hoa lng tnh, oi xng haiben.Baohoadangcanh,rinhau,xepthanh2vong:3manhvong ngoai(aihoa)va2manh vong trong(canhhoa)behnmanhth3 vongtrong.Manhnaycohnhdangvamausackhachan,goilacanh moi [13]. Qua: khi kho, m thanh 3-6 manh. Hat rat nho va nhieu, thng khong co noi nhu. Do nhe nen hat de phat tan nh gio. nhieu loai, trong qua co nhng long hut nc dung e ban hat i. Phoi trong hat phat trien yeu, khongphanhoathanhcquan.Hatmuonnaymamcanconamcong sinh [13]. 1.3ac iem ve phan loai. Orchidaceae la mot ho rat ln thuoc lp n t diep, phan bo khap ni tren the gii [16]. vung on i, ta gap nhieu loai song at nh a lan; mot so loai hoai sinhkhongdieplucvasongnhvaochatmucnattrongat;coloai Uc Chau co the song ngam di at nh nam. vung nhiet i, ta se gap nhieu loai phu sinh song tren cay khac nh Cattleya,Oncidium,LaeliataptrungnhieuvungTrungMy;ong NamAacsacnhatlaDendrobiumvaconcoCypripedium, Phalaenopsis, Cymbidium co nguon goc Indonesia. Mot so loai lan song tren a nh thach lan. Cay lan co the chia lam hai nhom [11]: Nhomnthan:ayla nhom ch tang trng ve chieu cao lam cay dai ra mai. Nhom n than chia lam hai nhom phu: -Nhomphulamocoi(Sarcanthinae):nhomnaylacxep thanh2hangmocoinhau,latrenmothangxenkevilacua hang kia. Gom cac giong nh: Vanda, Aerides, Phalaenopsis -Nhomphuladepthanghaytron(Campylocentrinae): Papilionanthe, Luisia Nhom a than: ay la nhom gom nhng cay tang trng lien tuc. Can c vao cach ra hoa nhom nay chia thanh 2 nhom phu: +~ .~+ :: ++- + .~+ :~ -. . .y-+ .. :+ ~+ y-+~+ 4 -Nhom ra hoa pha tren nh: Cymbidium, Dendrobium, Oncidium -Nhom ra hoa nh: Cattleya, Laelia, Epidendrum Ngoairaconcomotsogiongmangtnhchattrunggiannh: Centropetatum, Phachyphllum, Dichaea Hnh 2.1: Cau tao c quan sinh san. a.Cau tao hoa chi tiet. b.Qua lan chn. Hnh 2.2: Mot so giong hoa ep cua ho Orchidaceae. +~ .~+ :: ++- + .~+ :~ -. . .y-+ .. :+ ~+ y-+~+ 5 II.Gii thieu ve Dendrobium. 2.1Nguon goc va s phan bo: Giong lan nay c at ten vao nam 1799. Ch Dendrobium co nguon goc cua ch Hy Lap. Dendro co ngha la cay go, cay ln; bio la song, v tat ca cac loai cua Dendrobium eu la phu sinh song bam tren cay go [16]. Dendrobiumratphongphuvechungloai,naylnthnhcuahoLanvi khoang1.600loaiphanbotrencacvungthuocchauAnhieti,taptrungnhieu nhat ong Nam A va Uc chau [16]. ieu kien sinh thai cua Dendrobium cung rat a dang co nhieu loai ch moc va ra hoa vung lanh, co loai vung nong, co loai trung gian, va cung co loai thch nghi vi bat c ieu kien kh hau nao [11]. 2.2V tr phan loai: Lan Dendrobium thuoc: -Nganh:Angiospermatophyta -Lp: Liliopsida (Monocotyledones) -Lp phu: Liliidae -Bo: Orchidales -Ho: Orchidaceae -Giong: Dendrobium sp. Hnh 2.3: Tong quan ve lan Dendrobium. +~ .~+ :: ++- + .~+ :~ -. . .y-+ .. :+ ~+ y-+~+ 6 2.3ac iem ve hnh thai. Dendrobium la loai a than vi nhieu gia hanh, cac gia hanh thng mang mot than vi nhieu la moc xen ke, tren than co rat nhieu mat ngu. Can hanh vi khoang cach gia cac mat ngan hn Cattleya. Hoa co the moc t than thanh tng chum hay tng hoa co oc [11]. Hoa co mau trang, vang en tm. Thng la ai sau nam mot mnh, 2 la ai ben dai ra dnh lai vi nhau mep di va dnh vao ay cua tru tao thanh mot phan di chan cua tru pha di goi la cam. Moi gan vao cam, oi khi keo dai ve pha sau tao thanh ca, moc hay tui. Moi nguyen hay co thuy, gai, soc co long hay khong. 2 canh hoa ben giong nh 2 la ai. Tru thap. Phan c cua nh tru co nap ay, nap gan vao tru nh 1 ch ngan ve phai sau. 4 khoi phan nho dnh lai vi nhau tng cap, khong v, khong got. Dendrobium c chia ra lam 2 nhom theo dang than cua chung: -DangthonghayNobileladangthanmemthngvunghilanhnh a Lat. -DangnghayPhalaenopsis ladangthancngthngsongvungco kh hau nong hn. CaDendrobiumnobilevaDendrobiumphalaenopsiseucochungaciem trong viec tao lap cac gia hanh mi va trong s biet hoa choi s khi nach la doc theo gia hanh; nhng chung lai rat khac biet trong viec trong viec tao lap choi hoa. -Dendrobiumnobilerahoatchoiskhicuagiahanhatrng thanh. NhLong tu, Gia hac chung ch ra hoa vi gia hanh a rung het la. - Dendrobium phalaenopsis th hoa moc gia hanh cu lan gia hanh mi. giahanhmi,choinonnhatganngonlachoiautienphattrien thanh voi hoa. Hnh dang cua Dendrobium cung rat bien thien: -Nhom co gia hanh rat dai va mang la doc theo chieu dai cua gia hanh ay, thng rung het la khi ra hoa nh Long tu (Dendrobium primulinum), Y thao (Dendrobium gratio sissimum) -Nhom gia hanh to ngan , tan cung thng co 2-3 la dai, ben, khong rung. Phathoataptrungphannaytaothanhchumnghaythongnh: Thuytientrang(Dendrobiumfarmeri),Thuytienvang(Dendrobium thyrisflorum), Vay ca (Dendrobium lindleyi) -Nhom ca gia hanh rat manh mai, dai hay ngan, co la doc theo chieu dai cuachung,daibenkhongrung.HoathngcoocnachnhHng duyen (Dendrobium revolutum) Ngoai ra con co mot so loai Dendrobium khac cung thng c trong: -Kimiep(Dendrobium chysotosum var delacuorii):Hoavangti,moi vang di trung tam am. +~ .~+ :: ++- + .~+ :~ -. . .y-+ .. :+ ~+ y-+~+ 7 -Nhatiemhong(Dendrobiumdracoins):Hoatrangbongnhsapvi moi soc o ay. -Thach hoc (Dendrobium crumenatum): Hoa trang, moi co bt vang, thm nhng mau tan, t hoa nhng n ro cung luc. -Giahac(Dendrobiumanosmum,Dendrobiumsuperbum):Hoamau hng, co hai bt am mau trang hay tuyen. Rat thm [16]. 2.4Cac ieu kien c ban e nuoi trong lan Dendrobium sp.: 2.4.1Nhiet o. Nhietotacongcaylanquaconngquangtonghp,cngoquang hp gia tang theo nhiet o, thng nhiet o tang 10% th toc o quang hp tang len gap oi. Nhiet o con anh hng en s ra hoa mot so loai lan nh lan Bach cau Dendrobiumcrumenatumoihoigiamnhietokhoang5-60Ctrongvaigiayth9 ngay sau chung se n hoa ong loat. 18,50 Dendrobium nobile ch tang trng ma khong ra hoa nhng chung se ra hoa khi nhiet o ha xuong 130C hay thap hn [16]. Cancvaonhucaunhieto,cothetamchiaDendrobiumthanhhainhom chnh:-Nhom a lanh sinh trng va phat trien tot nhiet o ly tng la 150C, gom cac giong c lay t cac vung cao nguyen o cao tren 1.000m. cac loai nay neu c trong nhiet o cao hn hoac bang 250C, th cay van song, th cay van song nhng hiem khi ra hoa. -Nhomanong,nhietothchhpchocacloaicuanhomnayla250C, gom a so cac giong Dendrobium vung nhiet i, va cac loai cua giong Dendrobium lai hien ang trong tai thanh pho Ho Ch Minh va cac tnh pha Nam -Ngoaira,concomotnhomDendrobiumtrunggiancothesongca vunglanhvavungnong,nhngvunglanhcaysinhtrngvarahoa nhieu hn nh Dendrobium primulinum, Dendrobium farmeri nhiet o ly tng cua cac loai nay la 200C [11].2.4.2Am o. Caccaylan,nhatlaphonglan,songbamtrencaccaycao,chunglaynct cactranma,thinctrongkhongkh.Chnhamoquyetnhshiendie n cua cac loai phong lan [16]. Thong thng am o tng oi toi thieu 70% thch hp cho s tang trng cua nhieu loai. Tuy nhien am o ly tng van la am o cua vung ban x ma loai lan o c tm thay. Dendrobiumcungnhasocacgionglankhacchphattrientottrongkhong kh am va thoang. Am o tng oi can thiet la 40-70%. Cau tao gia the qua am va ung th bo re se b thoi va bieu hien la cay con keiki moc ra t phan ngon cua than [11]. +~ .~+ :: ++- + .~+ :~ -. . .y-+ .. :+ ~+ y-+~+ 8 2.4.3Anh sang. Anhsanglaieukiencanthietchossinhtrngvaphattriencuacaylan thong qua qua trnh quang hp. ay la yeu to quyet nh s tro hoa cua lan [16]. Dendrobiumlagiongasang,cothetrongtrongieukienanhsangtrctiep hay khuech tan. Anh sang hu hieu cho giong Dendrobiumla 70%. Neu tha anh sang, cay se b vang la, gia hanh b teo lai, cay xau i nhng cay sethchnghidan,vanrahoanhieuvaep.Neuthieuanhsang,caysebthoaihoa ro ret, cay eo uot va so lng hoa se t i [11]. 2.4.4Nhu cau phan bon. Dendrobiumthanngoihoidinhdngcao,chungcanratnhieuphanbon vacothedungnhieudangphanbonkhacnhau.ConcacloaiDendrobiumthan thong hap thu phan cham nen phai dung nong o that loang. Cac loai phan hu c nh: phan heo, banh dau kho, phan tom ca, phan trau bo kho co the dung rat tot bang cach pha loang vi nc roi ti, hoac vo chat tng vien at tren be mat gia the, re lan se hap thu dan dan cac dng chat c phong thch qua qua trnh ti nc. Cacloaiphanvoccdungthngcocongthc30-10-10dung3lan/tuan vi nong o 1muong ca phe/4lt. Trong suot mua tang trng, ta bon phan 10-20-30 lam 2 lan/tuan e tao mot sc chu ng cho cay trc khi bc vao mua ngh. Trong mua tang trng neu cay co nu hoa, ta thay phan 30-10-10 bang phan 10-20-20 vi chu ky bon nh tren cho en khi hoa tan. TrongmuanghhoantoankhongbonphanchoDendrobium,hayunghn giam va khong bon phan cho Dendrobium khi cay hoan tat thi k tang trng hang nam cua no. Khong nen dung cac loai phan rieng re, thng phan bon c dung dang hon hp va bo sung them cac chat phu gia la cac sinh to va cac nguyen to vi lng. 2.4.5Sau benh va cac van e khac. VlanDendrobiumcancbonnhieuloaiphanhuckhacnhauvamoi trng s da se muc nat sau mot thi gian ngan c trong. ay la 2 nguyen nhan gay ra nhieu sau benh hai. Mot loai rep dnh mau vang, kch thc rat be thng xuat hien tren be mat la. Loai nay gay tac hai tren cay qua viec hut nha. oi vi cac loai con trung can pha Dendrobiumth loai tr chung tng oi de dang bang Serpa, Bassa, nong o 1/500. Mac du Dendrobiumla cay khang benh rat manh, tuy nhien cay van b nam va virustancongneuieukienvesinhquakem.Nguyhiemnhatlabenhkhothan gan goc gia hanh do mot loai virus tham nhap lam cho cac gia hanh b kho va chet. CothengabenhbangcachnathangxtTopsil,Zineb,Benomylvinongo 1/400 [11]. +~ .~+ :: ++- + .~+ :~ -. . .y-+ .. :+ ~+ y-+~+ 9 Hnh 2.4: Mot so dang hoa ep cua giong Dendrobium. +~ .~+ :: ++- + .~+ :~ -. . .y-+ .. :+ ~+ y-+~+ 10III.Cac phng phap nhan giong. 3.1 Nhan giong ngoai thien nhien. 3.1.1 Nhan giong vo tnh. a.Tachbui:aylaphngphapaccacnghenhansdungoi vicacgionglanathannhCattleya,Dendrobium,Cymbidium, PaphiopeliumCattleya,Dendrobium,vanhnggiongtngt: moi goc cua gia hanh thng co t nhat mot mat ngu nen co the tach moi hanh gia thanh mot n v e trong. b.Chietcanh:Dendrobiumthngtaoracaycontrengiahanh (Keiki) mot cach t nhien. Khi cac cay con nay kha manh, co re tot, co the tach ra khoi gia hanh e trong. 3.1.2Nhan giong hu tnh. a.S thu phan: Trong thien nhien s thu phan lan do con trung thc hien. Cau truc cua hoa lan la hoan toan e thch ng cho s thu phan ay. Co hai phng phap thu phan: -Stthuphan:Khiphanhoacuabonghoanaycrivao nuomcuachnhhoaay.ieunayhiemkhixayratrongt nhienhoalanvcautruccuabophansinhduccvacai hoa lan. -Sthuphancheo:Khiphanhoahoanaycevaonuom cuahoakhaccuacungcayhaycungloai(thngxayratrong thien nhien do con trung thc hien), hoac khac loai, khac giong (thng do con ngi thc hien). b.Qua lan: Neu s thu phan co ket qua, th co the ngay trong ngay hay sangngayhomsau,cacphienhoaxulainhngkhongrung.Vae tranhsthuphankhacdocontrungngitadungbaonilongtrum hoa lai, nhng khong buoc kn mieng v ham hi se lam h trai. Sau khi thu phan, bau noan t t trng phu to ra thanh trai. Moi trai co the cha hang ngan hay en ca trieu hot. Khi tra t mau xanh luc chuyen sang mau vang luc th nen hai trai. c.Gieo hat: Trongthiennhienmuonhatlannaymamthhatlanphaic nhiemmotloainamksinh.Ngitaakhampharamotsoloai namgiupnaymamhatlan,moiloaichgiupnaymammotso giong lan ma thoi.-RhizoctoniarepensgiupnaymamCattleya,Laelia, Angraecum, Cypripedium (Paphiopedilum). Download http://Agriviet.Com+~ .~+ :: ++- + .~+ :~ -. . .y-+ .. :+ ~+ y-+~+ 11-Rhizoctonia mucoroides giup nay mam Vanda, Phalaenopsis. -RhizoctonialanugiosagiupchohotnaymamOncidium, Odontoglossum va Miltonoa [16]. 3.2Nhan giong trong phong th nghiem. TrongphongthnghiemlanDendrobiumcchungtoinhangiongbang phngphapgieohatin-vitro.Trongthiennhien,vhotlanquanho,khongcha chatdtrvachcomotphoichaphanhoa,nenkhongthephattrientheomot phngcachbnhthngcvavvaymaviecchohotlannaymamvaphat trien thanh cay lan trng thanh la van e kho khan. Ngi trong lan a tm nhieu cachgieohatnhngkhongthanhcong.Nam1922,KnudsonMy,thanhcong trongviecthaythenambangngmoitrngthachegieohat[16].Phng phap nay co nhieu u iem nh: -Kethpvikythuatlaigiongtaoranhieugionglanmi cung cap cho th trng. -Giupbaotoncacloailanquy,vanhannhanhsolnggiup cho cong tac bao ton. IV.S lc ve ky thuat nuoi cay mo thc vat. Nhangionginvitrohaynuoicaymoeulathuatngmotacacphngthc nuoicaycacbophanthcvattrongongnghiemcochamoitrngxacnh ieukienvotrung[5].Kythuatinvitrodatrennguyenlylatebaothcvatco tnh toan the, ngha la t mot mo, mot c quan hoac mot te bao cua bat ky bo phan naocuacayeucothephattrienthanhmotcayhoanchnhneucnuoitrong moi trng thch hp [6]. 4.1ieu kien nuoi cay te bao thc vat. a.Yeucaucbannhatcuaphongnuoicaymolaphaiambaovo trung. Khai niem vo trung nay bao gom vo trung moi trng nuoi cay va ca s bao am sao cho mau nuoi cay c hoan toan vo trung. b.Cac yeu cau c ban cua ky thuat nuoi cay mo te bao thc vat: -Khi thiet lap phong nuoi cay mo thc vat phai am bao c tnhlientucthuanlichocacthaotac,cacgiaioantrong suot qua trnh nuoi cay mo. -ambaocvesinh(tnhvotrung)cuasanphamcuoi cung. -Chuanbmoitrngungcach,chonungmoitrngcho tng loai thc vat va tng giai oan nuoi cay. -Chon va x ly mo thch hp trc khi cay [7]. Download http://Agriviet.Com+~ .~+ :: ++- + .~+ :~ -. . .y-+ .. :+ ~+ y-+~+ 12c.Cacthanhphancbancuamoitrngnuoicaybaogom:nc, muoikhoang(alngvavilng),nguoncacbon,vitamin,chat ieu hoa sinh trng, agar (oi vi moi trng ran) [14]. -Nc: nc dung trong moi trng nuoi cay la nc cat, hoac nc kh ion, tot nhat la nc cat hai lan t cac may cat nc hoan toan bang thuy tinh. -ng: tao nguon cacbon giup mo tong hp nen cacbohydrat chocaymakhongphaidoquanhhpcuamocaytaonen; ng s dung pho bien la saccarose (1-6%w/v), glucose (2% w/v). -Cac muoi khoang a lng: nuo i cay mo thc vat oi hoi mot lng nhat nh cac nguyen to a lng nh N, P, K, Ca, Mg, S, Fe, -Cacmuoikhoangvilngcovaitroquantrongtrongi songthcvat.Chungthamgiavaocacquatrnhsinhhoa trong te bao, tham gia vao trung tam hoat tnh cua enzyme va vitamin,tangtnhchongchucuathcvatvicacieukien moi trng bat li. Cac vi lng thong dung la Mn, B, Zn, Cu, Co, I, nong o s dung trong moi trng rat thap. -Vitamins: chu yeu myo-inositol, acid nicotinic, puridoxin HCl (B6), thiamin HCl (B1)Cac vitamin de b hong do nhiem tao nen can gi t0< 00C [9]. -Chatieuhoatangtrng:tuytheotngmucchnuoicay co the chon cac nong o va to hp cac nong o va to hp cac chatieuhoasinhtrngphuhp.Cacchatieuhoasinh trngthngdunglaIAA,NAA,2,4D,Kinetin,BAP, IBA[14] -Agar(oivimoitrngac):lamotpolisacaritlamtrong bien.Agartan1000Cvakhinguoiseongaclai.Ham lng s dung t 8-10g/l tuy hang che tao[10]. d.Nhiet o, anh sang va pH anh hng en qua trnh cay: -Nhieto:nhietocuaphongnuoicaythngcieu chnh on nh t 22-250C. moi te bao thc vat khac nhau th nhiet o nuoi cay cung khac nhau [5]. -Anh sang: co anh hng en qua trnh phat sinh hnh thai cay nuoi cay. Cac yeu to anh hng bao gom: cng o, chu ky, thanh phan quang pho anh sang. Cng o anh sang t: 1000-2500 lux c dung pho bien cho nuoi cay nhieu loai mo [5]. -pH:lamotyeutoquantrong.SonnhpHmoitrngla yeutoduytrtraooicacchattrongtebao.eieuchnh Download http://Agriviet.Com+~ .~+ :: ++- + .~+ :~ -. . .y-+ .. :+ ~+ y-+~+ 13pHmoitrngcothedungdungdch10%hoac1NNaOH hoac 1N HCl [14]. 4.2 Ky thuat gieo hat trong ong nghiem. a.ac iem cua hat lan. Khonggiongnhngloaihatkhac,hatlanratnho,nhhatbuiva khongchanguondtrthcan.Trongtnhien,hatcothenay mamnhngsekhong phat trien neukhongcostacongcuanam mycorrhizal [22]. Loai nam nay tieu hoa cac vat chat hu c tren cay chuhoactrongat,chuyenhoathanhcacngngiangiuphat lan nay mam va giup phoi phat trien [23]. Khi hat lan a nay mam, nosetaoramotlnglncactebaokhongphanhoacgoila protocorm. Neu tat ca eu tot, protocorm se tiep tuc trongvai tuan, vai thang, tham ch ca nam tuy loa i, cho en khi u ln e tao than va re. cac loai lan at, duy tr moi quan he cong sinh nay rat quan trongtrongsuotgiaioanautiencuacay,vprotocormnamdi atnenkhongthettonghpthcan.cacloailanphusinh, protocorm thng xanh, va v the no co the t tong hp c mot t thc an [22]. b.Kh trung trai lan. Viec kh rung trai lan co the thc hien theo hai phng phap: -Qualansaukhimora,hatthuctrckhiemgieocan tuyen chon cac hat chac, cac hat lep khong thu tinh can phai loaiboibangcachemlytamtrongnccat;cachatlan totselangphadiongnghiem,hatlepnoilentren,c ganboi.Cachattotctieptuckhtrungbang Hypochlorite de calcium 10%. Sau ay, cho hat lan va nc kh trung vao ong nghiem, va uphuknhatlan,lacmanh,euvaeyentrongvong10 phut,sauoonckhtrungrabangonghut,vatieptuc ra hat lan 3 lan bang nc cat vo trung. Cuoi cung hat lan a kh trung se c gieo vao cac bnh cha moi trng. -Khi qua lan a c cat tren cay, mang vao phong th nghiem, can e ni sach se, thoang mat. Neu cha gieo hat ngay th co the gi qua trong tu lanh, trong vong 3-4 ngay. Khigieohat,trctienqualancancatbocaccuongdai, uoiconsot lai cac la ai, canh hoaDung bong gon co tam alcol 700 e lau toan bo mat ngoai cua qua. Mot khi vao tu cay vo trung, qua lan se c: -Nhung vao alcol 90-960, trong 1 phut, h nhanh qua ngon la. -Nhung vao Hypochlorite de calcium 10%, trong 10 phut, ra lai 3 lan bang nc cat vo trung. Download http://Agriviet.Com+~ .~+ :: ++- + .~+ :~ -. . .y-+ .. :+ ~+ y-+~+ 14Sau ay qua lan se c cat got hai au, va dung dao mo xe doc qua lan (mo hai ng oi dien nhau), tach lam hai qua lan, ke en dung pincevadaomoe gi lay tatca cac hat co trong hai na qua lan, bang cach go nhe len phan ngoai cua vo qua. Neu s thu tinh tot, tat cacachatsecomauvangvaeurixuongmotapetridunge hng lay hat. Sau o ngi ta se dung muong hoac mot dung cu inox dung e gieo hat vao cac bnh cha moi trng [8]. c.Kythuatgieohattrongongnghiem:hatsenaymamtrongong nghiembangthuytinhhaybangnhatrenmoitrngrancocha ngvacacloaichatkhoangcanthietehatnaymamvaphat trien.oivilancohaikieunaymamcban:naymamcannam cong sinh va nay mam khong cong sinh. -Trong kieu nay mam cong sinh, hat c gieo vi mot lng nhonammycorrhizalthchhp.Namnaysephattrientren moitrng,baophucachatnaymamvamoiquanhecong sinh xuat hien se duy tr sc song cua protocorm cho en khi notaoralavacokhanangtdng.Kythuatnaycs dungrongraienhangiongcacloailanatvungoni. Kythuatnaycolithelamoitrngsdungratngian (mottrongnhngdangphobienchbaogombotyenmach vamottdchchietnammen),vacaysinhrathngkhoe hnvacokhanangekhangvicactacongcuanamtot hn cay oi chng nay mam khong can nam. Ky thuat nay co motnhciemlacanphaicomotchungnammycorrhizal thuankhiet,hoacquanhecongsinhkhongphattrien,hoac trthanhkysinhvahatgieosechet.Corattnghiencu thc hien ve cac loai nam mycorrhizal cac loai lan nhiet i, nen ky thuat nay t c ap dung. -Kieu nay mam khong cong sinh thng c s dung e nhan giongcacloailannhieti.Moitrngsdungphctap hn bao gom cac muoi khoang, cac chat hu c va ng vi mot lng thch hp e hat lan nay mam ma khong can nam [22]. d.ng dung cua viec gieo hat. -Gia tang lng hoa lan tren th trng -Nhangiongcacloailanhiemnhannhannhanhsolngca trong t nhien va trong cac chng trnh bao ton. -Cungcapcayconeban,giuphancheskhaithaccay hoangdai,kchthchsquantamcuacongchungvatao nguon thu ngan sach. -Kethpvikythuatlaigiongtaoranhngcaylankhoe manh co mau sac ac biet [22]. Download http://Agriviet.Com+~ .~+ :: ++- + .~+ :~ -. . .y-+ .. :+ ~+ y-+~+ 15V.Cac cong trnh ngien cu gieo hat lan trong ong nghiem. 5.1 Trong nc: DngCongKienvacongsnghienculaitaovagieohatlan Dendrobium trong ong nghiem [8]. 5.2 Ngoai nc: Nam1844,Neumann,motngiPhaplamvn,alamnaymammotsohot lan bang cach rai ai cac hot lan tren cac cuc at quanh goc cua cac cay lan ln. Nam 1899, Noel Bernard, nha thc vat ngi Phap, a kham pha ra b mat cua viecnaymamhotlankhiongkhaosatcachotNeottianidus-avisnaymamt nhien trong rng. Nam 1904, ong cong tac vi Burgeff e a ra phng phap gieo hot lan co nhiem nam trong chai thach [16]. Nam 1909, Hans Burgeff a lam nay mam c Laeliocattleya tren moi trng dinh dng gom 0,33% saccarose (ng ma). Nam 1922, Lewis Knudson a lam nay mam Cattleya mossiae tren moi trng Pfefferds 1% saccarose (ng ma) [11]. VI.Muc ch gieo hat lan Dendrobium trong ong nghiem. Hnh 2.5: Qua lan Dendrobium. Cac nha trong lan khong ngng tm kiem cac giong lan mi e thoa man s hieu k cua mnh hoac co the em ban. Co hai cach e co c giong lanmi.Motla,sutapnhnggionglanhoangdaitrongrngemve thuan hoa, tao cac ieu kien nhan tao giong t nhien e cay lan co the ra hoa.Phngphapnaygapnhieuruirodoieukienmoitrngkhong thuan li cho cay lan phat trien. Haila, tao ra nhng giong lan lai mi, caylanlaisemangnhngactnhtotvttroicuacabome,cothe thoa man c nhu cau cua ngi thng thc lan. Tuy nhien khi hai cay lan lai vi nhau at ket qua va tao trai can phai ket hp vi phng phap gieo hat trong ong nghiem e hat lan co the nay mam de dang. Co nh vay mi co the kiem tra ket qua cua viec lai hai cay lan. oi vi mot so giong lan quy can phai bao ve th phng phap gieo hat trong ong nghiem giup bao ton giong, nhan nhanh so lng cay ma van am bao c tnh on nh di truyen. Download http://Agriviet.Com+~ .~+ :: ++- .~: -. +++ +~ . .y-+ .. :+ ~+ y-+~+16 PHAN III: VAT LIEU PHNG PHAP I.Vat lieu va ieu kien gieo hat trong ong nghiem. 1.1 Mau qua gieo hat. Vat lieu la qua lan Dendrobium lai dai t 4-5 cm lay t cay ngoai thien nhien, tai vn thay Dng Cong Kien, quan 12, thanh pho Ho Ch Minh. 1.2 Moi trng nuoi cay. Moi trng Knudson C (1946), bo sung them: -Nc da 15%. -Than hoat tnh 0.5g/l, tuy theo muc ch th nghiem. -ng 30g/l. -Agar 8,5 g/l. -pH = 5.7 6. Moi trng MS (Murashige va Skoog, 1962), bo sung them: -Nc da 15%. -Than hoat tnh 0.5g/l, tuy theo muc ch th nghiem. -Glycin 0.5g/l -myo-Inositol 0.1mg/l -ng 30g/l. -Agar 8,5 g/l. -pH = 5.5 5.7. S dung chai thuy tinh 100 ml va 500 ml e nuoi cay. Tat ca moi trng c hap kh trung 1atm, 1210C trong thi gian 20 phut. 1.3 ieu kien nuoi cay. Thnghiemcthchientngay20/01/2005enngay26/05/2005,taicong ty Van Thanh Tai, 114 Hong Ha, Quan Tan Bnh, Thanh pho Ho Ch Minh, vi cac ieu kien nuoi cay nh sau: -Nhiet o phong nuoi: 24-280C. -Cng o chieu sang: 2000-3000 lux. -Thi gian chieu sang: 8 gi/ngay. -Am o trung bnh: 60-70%. Download http://Agriviet.Com+~ .~+ :: ++- .~: -. +++ +~ . .y-+ .. :+ ~+ y-+~+17 II.Hng i cua e tai. oi vi e tai, chung toi hng muc ch nhan giong lan Dendrobium lai bang phng phap gieo hat in-vitro nham: -Cung cap giong lan mi cho cac phong nuoi cay mo. -Tao ra cay con khoe, sach benh, e trong ra vn m. -Kiem tra ket qua lai. Vi thi gian qua ngan, t nguyen lieu ban au la qua lan, chung toi ch co the thc hien c cac giai oan: -Kh trung qua. -Nhan protocorm. -Ra re. III.Bo tr th nghiem: 3.1Th nghiem 1: Kh trung qua a.Muc ch th nghiem: -Tm phng phap kh trung thch hp. -TmnongoHypochloridecalciumCa(OCl)2vathigiankh trung thch hp oi vi qua lan. -Theo doi bieu hien cua hat. b.Phng phap va hoa chat s dung: -Phng phap ot con. -Phng phap hoa chat: dung Ca(OCl)2. c.Bo tr th nghiem: -Mau lay t thien nhien. -Ra mau di nc chay khoang 10 phut. -Ra mau bang xa phong pha loang. -Ra lai nhieu lan bang nc cat. Chuyen mau vao tu cay vo trung. -Ra mau bang con 700 trong 1-2 phut. -Ra mau lai bang nc cat vo trung khoang 2-3 lan. -Ngam mau trong Ca(OCl)2 va thng xuyen lac mau. -Ralaimaunhieulan bangnccatvotrung khoang3-4lanchothat sach. -Nhung vao con 900 va h nhanh qua ngon la. -Nhanhchongatqualanvao a petri vao ay nap lai. Download http://Agriviet.Com+~ .~+ :: ++- .~: -. +++ +~ . .y-+ .. :+ ~+ y-+~+18 -Sau ay qua lan se c cat go t 2 au, va dung dao mo xe doc qua lan, tach lam 2. -Dung dao gi phan vo qua, dung kep lay het hot ra mot a petri. -Gieo hatvao moi trng. -Th nghiem c lap lai 2 lan, moi lan mot qua. d.Ch tieu theo doi. Ty le hat song sau khi gieo. Tnh trang hat gieo:Xanh. Trang. Hoa nau. Download http://Agriviet.Com+~ .~+ :: ++- .~: -. +++ +~ . .y-+ .. :+ ~+ y-+~+19 Hnh 3.1: Quy trnh kh mau. Download http://Agriviet.Com+~ .~+ :: ++- .~: -. +++ +~ . .y-+ .. :+ ~+ y-+~+20 3.2Th nghiem 2: Tm moi trng thch hp e gieo hat. a.Muc ch th nghiem. Tm moi trng khoang thch hp e gieo hat. b.Moi trng gieo hat. -Moi trng MS khong bo sung kch thch to, ky hieu Mo. -Moi trng MS bo sung 1ppm BA, ky hieu la M1. -Moi trng KnudsonC khong bo sung kch to ky hieu Ko. -Moi trng KnudsonC bo sung 1ppm BA, ky hieu K1. c.Bo tr th nghiem. -Cacqualanackhtrungtthnghiem1,secgieo vao 4 loai moi trng th nghiem. -Theo doi va ghi nhan ket qua sau 2-4 tuan -Th nghiem c lap lai 2 lan , moi lan 15 chai 100ml. d.Ch tieu theo doi. -Thi gian hat nay mam. -Moi trng thch hp gieo hat Dendrobium. Download http://Agriviet.Com+~ .~+ :: ++- .~: -. +++ +~ . .y-+ .. :+ ~+ y-+~+21 3.3Th nghiem 3: Tao Protocorm a.Muc ch th nghiem. -So sanh s thanh lap protocorm tren hai moi trng khac nhau va bo sung cung nong o kch thch to. -Tm moi trng thch hp cho cac th nghiem ve sau. b.Moi trng nuoi cay. -Moi trng MS khong bo sung kch thch to, ky hieu Mo. -Moi trng MS bo sung 2ppm BA, ky hieu la M2. -Moi trng KnudsonC khong bo sung kch to ky hieu Ko. -Moi trng KnudsonC bo sung 2ppm BA, ky hieu K2. c.Bo tr th nghiem. -Nhng mau hat a phnh to ra, co mau xanh, khong nhiem t th nghiem 2. -Theo doi va ghi nhan ket qua sau 2-4 tuan nuoi cay. -Th nghiem c lap lai 3 lan, moi lan 8 chai 100ml. d.Ch tieu theo doi -Thi gian tao protocorm. -Moi trng tao protocorm tot nhat.-Tnh trang phat trien cua prorocorm. Download http://Agriviet.Com+~ .~+ :: ++- .~: -. +++ +~ . .y-+ .. :+ ~+ y-+~+22 3.4Th nghiem 4: Nhan protocorm a.Muc ch th nghiem. So sanh s nhan protocorm tren cung mot loai moi trng vi nong o kch thch to thay oi. b.Moi trng nuoi cay. -MoitrngKnudsonCoichngkhongbosungkchthchto ky hieu K0. -MoitrngKnudsonCbosungBAvinongo 1ppm:2ppm:3ppm, ky hieu la K1, K2, K3. -MoitrngKnudsonCbosung2ppmBAvaIAAvinongo 0.1ppm:0.5ppm:1ppm, ky hieu la K4, K5, K6. c.Bo tr th nghiem. -S dung cac protocorm c tao thanh t th nghiem 3. -Theo doi va ghi nhan ket qua sau 2-4 tuan nuoi cay. -Th nghiem c lap lai 3 lan, moi lan 3 chai 500ml. d.Ch tieu theo doi. -Nhanxetmoitrngnhannhanhprotocormbatchoitotnhat: thi gian ngan nhat cho cay co chieu cao 2-3 cm va co 2-3 la. -S tao thanh cay gia cac moi trng.-Mau sac va chieu dai la. -Thi gian xuat hien re au tien. Download http://Agriviet.Com+~ .~+ :: ++- .~: -. +++ +~ . .y-+ .. :+ ~+ y-+~+23 3.5Th nghiem 5: Ra re a. Muc ch th nghiem. Khao sat moi trng thch hp cho s ra re. b. Moi trng nuoi cay. -MoitrngKnudsonCoichngkhongbosungkchthchto ky hieu K0. -MoitrngKnudsonCbosungNAAvinongo 0.1ppm:0.5ppm:1ppm, ky hieu la KN1, KN2, KN3. -MoitrngKnudsonCbosungIAAvinongo 0.1ppm:0.5ppm:1ppm, ky hieu la KI1, KI2, KI3. -MoitrngKnudsonCbosungIBAvinongo 0.1ppm:0.5ppm:1ppm, ky hieu la KB1, KB2, KB3. c. Bo tr th nghiem. -Nhngcayconcochieucaot2-3cmvaco2-3lactao thanh t th nghiem 4. -Theo doi va ghi nhan ket qua sau 3-4 tuan nuoi cay.-Th nghiem c lap lai 3 lan, moi lan 3 chai 500 ml. d. Ch tieu theo doi -Chieu dai re. -S tang trng chieu cao cay. Download http://Agriviet.Com+~ .~+ :: ++- -: .~ +~ .~+ . .y-+ .. :+ ~+ y-+ ~+ 30 PHAN IV: KET QUA THAO LUAN I.Th nghiem 1: Kh trung mau. Qua lan sau khi c kh trung, gieo hat vao moi trng nuoi cay, sau 1 tuan ket qua ghi nhan c nh sau: Bang 1: Ket qua kh trung mau bang Ca(OCl)2, sau 1 tuan. Song va khong nhiemChet va khong nhiemNhiemNTSo lan ot So lngTy le (%) So lngTy le (%) So lng Ty le (%) 11161000000 221062,5637,500 Bang 2: Ket qua kh trung mau bang phng phap ot, sau 1 tuan. Mau hat lan c gieo vao moi trng, sau 1 tuan kh trung, mau bat au co nhng bieu hien khac nhau: -oivinhngmausongvakhongnhiem,sau1tuan,hatdan danthchnghivimoitrng,batauchuyensangmaunau vang mat ong. -Con vi nhng mau hat b anh hng bi Ca(OCl)2, va nhiet o cao, hat chuyen sang mau trang va nau uc, khong phat trien. Ket qua ghi nhan c bang 1 cho thay: -Nongo Ca(OCl)2 7%, thi gian kh trung la 15 phut ty le hat songvakhongnhiemla75%,maukhongbchetnhngtyle Song va khong nhiemChet va khong nhiemNhiemNT Nong o Ca(OCl)2 (%) Thi gian (phut) So lngTy le (%) So lngTy le (%) So lngTy le (%) 1715127500425 27201593,7516,2500 3915161000000 49201062,5637,500 Download http://Agriviet.Com+~ .~+ :: ++- -: .~ +~ .~+ . .y-+ .. :+ ~+ y-+ ~+ 31 nhiem cao (25%). Con vi thi gian kh trung 20 phut, ty le hat song kha cao (93,75%), mau hoan toan khong b nhiem, nhng b chet (6,25%). -Khi kh trung nong o tang th ty le mau b chet tang theo va so mau nhiem giam dan. nong o 9%, thi gian kh trung la 15 phut, ty le hat song rat cao (100%), mau hat gieo hoan toan khong b nhiem, khong chet. Khi keo dai thi gian kh trung 20 phut, th ty le mau chet tang cao (37,5%). Khi ngam qua lan trong dung dch Ca(OCl)2, phan vo ben ngoai tiep xuc trc tiep vi dung dch kh trung, con phan hat ben trong khong b anh hng. Khi gieo hat vao moi trng, hat co kha nang phuc hoi nhanhvanaymam.Tuynhien,neutangnongovakeodaithi gian kh trung, dung dch se ngam vao ben trong vo qua, nen sau khi gieo vao moi trng, hat phuc hoi rat cham va co the se b chet (hoa trang).Nhvay,vicacnongoCa(OCl)2 acsdungtrongloatth nghiem, th nong o 9% trong thi gian 15 phut la thch hp nhat. Ket qua ghi nhan c bang 2 cho thay: -Khtrungqualanbangphngphapotconthmauhatgieo hoan toan khong b nhiem. Nhng neu tien hanh ot 2 lan th ty lemauchetcao(37,5%),tngngviphngphapkh trung bang Ca(OCl)2 9%, trong thi gian 20 phut.-Khi ot mot lan ty le mau song la 100%, tng ng vi khi kh trung bang Ca(OCl)2 9% trong thi gian 15phut. Khi nhung qua lan vao con 900 va ot, th phan vo qua ben ngoai se banhhngtrctiepcuanhietocao,ongthiseloaibohoan toan cac tac nhan gay nhiem ben ngoai vo. Nhng neu tien hanh ot 2 lan, nhiet o cao cung vi thi gian keo dai se gay anh hng en hatbentrong,nenkhigieovaomoitrng,mauhatphuchoirat cham, hoac co the hoa nau, khong nay mam c. Nhvay,khikhtrungqualanbangCa(OCl)29%trongthigian15 phut va ot con mot lan, ty le song, khong nhiem la nh nhau. Tuy nhien, dung phng phap ot con th hat gieo phuc hoi va nay mam nhanh hn. Ngoai ra, phng phap nay con tiet kiem nhieu thi gian kh trung mau, thao tac n gian hn, kinh te hn. Tom lai: mau kh trung la qua lan Dendrobium con xanh. V mauhat gieo nam bentrong, nenneutienhanhra mau ky vaotbangcon1lanthcotheloaibo hoantoancactacnhangaynhiem.Hatkhongbanhhngvakhigieovaomoi trng se phuc hoi rat nhanh. Download http://Agriviet.Com+~ .~+ :: ++- -: .~ +~ .~+ . .y-+ .. :+ ~+ y-+ ~+ 32 Download http://Agriviet.Com+~ .~+ :: ++- -: .~ +~ .~+ . .y-+ .. :+ ~+ y-+ ~+ 33 II.Th nghiem 2: Cac qua lan sau khi c kh trung se c gieo vao 4 loai moi trng khac nhau, c theo doi sau 4 tuan nuoi cay: Moi trngKy hieuNong o BA (ppm) So mau hoa nau(1 tuan)% So maunay mam (2 tuan)% So mauhoa xanh (4 tuan)% M0 010083,3393,33MS M1 110090100 K001005063,33KC K1 110043,3380 Bang 3: S nay mam cua hat lan tren moi trng KnudsonC va MS bo sung 1 ppm BA. -Mottuansaukhigieovaomoitrng,tatcamauhatlaneu hoa nau vang. -Sau 2 tuan, mau bat au co hien tng nay mam va co s khac biet ro ret gia cac loai moi trng khac nhau.-Sau 4 tuan, nhng hat co mau hi vang, dan chuyen sang xanh do co s tiep xuc vi anh sang, tao c diep luc to. Da vao bang ket qua, chung toi nhan thay: -HatgieotrongmoitrngMScotylenaymamratcao(80-90%),haunhtatcacacmauhateucohientngphnhlen vachuyensangvang.Trongkhio,mauhatgieotrongmoi trng KnudsonC, ty le nay mam rat thap (40-50%). Trong thi iem nay hau nh khong co s khac biet gia loai moi trng co va khong co kch thch to. Co the trong thi iem nay, kch thch tokhongcoanhhnggmayoivikhanangnaymamcua hat. -Sau 4 tuan, hat lan hoa xanh. Trong moi trng MS co bo sung BA 1ppm, tat ca mau hat eu hoa xanh. neu khong co kch thch totylenayla93,33%.TrongmoitrngKC,tylemauhoa xanh thap hn, ay co s khac biet ro ret gia moi trng co va khong co kch thch to. Moi trng KC co BA ty le nay mam caohn(80%)sovikhongcoBA(63,33%).Tronggiaioan nay, vai tro cua kch thch to a co bieu hien ro ret. Cac mau hat Download http://Agriviet.Com+~ .~+ :: ++- -: .~ +~ .~+ . .y-+ .. :+ ~+ y-+ ~+ 34 gieotrongmoitrngcokchthchto,comauxanhamhn, hat phnh to hn T cac ket qua ghi nhan c cho thay, cung nong o kch thch to, nhng tren moi trng MS co nong o khoang cao hn, th hat lan nay mam nhanh hn so vi moi trng KnudsonC. Vay, moi trng MS co 1 ppm BA cho s nay mam cua hat lan Dendrobium tot hn so vi moi trng KnudsonC. Download http://Agriviet.Com+~ .~+ :: ++- -: .~ +~ .~+ . .y-+ .. :+ ~+ y-+ ~+ 35 III.Th nghiem 3:Cac mau hat lan sau khi a hoa xanh, c cay chuyen qua cac moi trng khac nhau e tao protocorm, c theo doi sau 4 tuan nuoi cay: Tnh trang tao protocormMoi trngKy hieuNong o BA (ppm) Sau 2 tuanSau 4 tuan M0 00+MS M2 2++ K00+++KC K2 2+++++ Bang 4: Ket qua tao protocorm tren moi trng KnudsonC va MS bo sung 2 ppm BA. Vi0: khong co s tao protocorm. + : tao thanh protocorm rat t. ++ : tao thanh protocorm nhieu. +++ : tao thanh protocorm rat nhieu. -Tuan au tien tren ca 4 moi trng eu khong tao protocorm. -Sau 2 tuan, hat gieo bat au bien oi tao thanh mot am te bao co mau xanh. -Solngprotocormngaycangtangnhanhsau3-4tuan,phnh to len, va co la mam. Da vao bang ket qua chung toi nhan thay: -Hai tuan sau khi cay, moi trng MS, hat lan khong co s bien oiroret,taothanhrattprotocorm.Saubontuan,solng protocorm tao thanh nhieu, ng knh nho, mau xanh hi trang, la mam dai khoang 3-4 mm va rat t . -TrongkhiomoitrngKnudsonC,sauhaituantatcacac hat lan eu bien oi ro ret, ng knh protocorm khoang 2mm, trenprotocormcolonghut.Sau4tuan,solngprotocorm trongmoitrngKnudsonCphattriennhanh,mauxanh,la mam dai khoang 5-7 mm. Download http://Agriviet.Com+~ .~+ :: ++- -: .~ +~ .~+ . .y-+ .. :+ ~+ y-+ ~+ 36 -TrongmoitrngKnudsonCcochaBA,macdusolng protocormtaoratngngvimoitrngkhongchaBA. Nhng protocorm phat trien tng oi nhanh hn, s tang trng cua protocorm ong eu vi nhau, co nhieu la mam khoe, mau xanhti.TrongmoitrngcochakchthchtoBA,lamot cytokinincokhanangkchthchvaieuchnhsphanbao, tang s chuyen hoa chat dinh dng [10].Tom lai:-Tnhngketquaghinhancvethigianvasolng protocormtaothanhtacotheketluanrang:staoprotocorm trong moi trng KnudsonC co bo sung kch thch to BA la tot nhat.-Do s tao thanh protocorm trong moi trng KnudsonC tot hn, nentatcacacthnghiemvesau,chungtoichthchientren moi trng KnudsonC. Download http://Agriviet.Com+~ .~+ :: ++- -: .~ +~ .~+ . .y-+ .. :+ ~+ y-+ ~+ 37 Download http://Agriviet.Com+~ .~+ :: ++- -: .~ +~ .~+ . .y-+ .. :+ ~+ y-+ ~+ 38 IV.Th nghiem 4: Cac protocorm c tao thanh t hat co chat lng nh nhau c a vao cac th nghiem nhan protocorm, sau 4 tuan theo doi ket qua ghi nhan c nh sau: Kch thch toNong o (ppm) Ky hieu So lng choi mi Chieu cao choi (cm) Chieu dai la (cm) Ghi chu 00K0000Protocorm nho, t. 1K11-21,5-20,5-0,7Protocorm mi xanh, nhng t. 2K24-61,5-2,50,5-0,7Protocorm nhieu, to, xanh, co long hut. BA 3K31-21,3-1,50,5-0,7Protocorm nhieu nhng nho. 0,1K44-62-3,51,5-2Protocorm xanh, to, co long hut, nhng t 0,5K55-73-42-3Protocorm xanh, nhng t 2ppm BA + IBA 1K64-52-31,5-2,5Protocorm nho, hi vang, t Bang 5: Ket qua nhan protocorm tren moi trng KnudsonC vi kch thch to khac nhau. -Sau2tuan,batauxuathienprotocormmi,colonghut.So lng va kch thc thay oi tuy theo moi trng. -Sau 3 tuan, co mot so protocorm bat choi mi. -Sau4tuancacchoimiphattrienmanh,tuytheomoitrng ma so lng va chieu cao choi mi khac nhau. Da vao bang ket qua chung toi nhan thay: -Sau2tuan,moitrngK1,khongthaysxuathiencua protocormmi.Sangentuanth3,mixuathienprotocorm mi. So lng protocorm mi tao thanh rat t, co mau xanh. Mot so protocorm bat choi vao giatuan th 4, nhng so lng nay rat t, ch khoang 1-2 choi. -moitrngK2,snhanprotocormdienranhanhvamanh. Sau2tuan,axuathiennhieuprotocormmi.Sau4tuan,so lngprotocormmitaothanhratnhieu,cacprotocormnay xanh mt, to, xung quanh co nhieu long t trang mn. Vao gia Download http://Agriviet.Com+~ .~+ :: ++- -: .~ +~ .~+ . .y-+ .. :+ ~+ y-+ ~+ 39 tuan th 3, co mot so protocorm mi bat choi, tuy so lng choi nhieu, nhng chieu cao choi rat thap. -So lng protocorm mi tao thanh nhieu moi trng K3, sau 2 tuan,axuathiennhngprotocormmi.Sangentuan4,so lng protocorm a rat nhieu, tuy nhien cac protocorm nay nho, co mot so protocorm co mau trang. Hau het cac protocorm mi eu khong bat choi. Trong cac moi trng K1, K2, K3, di tac dung cua kch thch to BA, cac mau cay eu tao thanh protocorm mi. Cac protocorm tao thanh cangnhieukhinongokchthchtotanglen,nhngchatlng protocorm th ngc lai (protocorm nho va trang). Nh vay, vi nong okchthchtothchhpsetacongtotensphanchiatebao, nhng khi nong o kha cao, th cung co the gay oc. -Sau2tuan,trongcacmoitrngK4, K5, K6,athayxuathien protocormmi.Tuynhien,sau4tuan,solngprotocormnay haunhkhongoi.TrongmoitrngK5vaK6,cacprotocorm comauxanhmt,to,colonghut.Solngprotocormtrong moi trng K6, t hn, protocorm co mau xanh hi vang, nho. -Sang en tuan th 3, cac protocorm bat choi, sang en tuan th4 cac protocorm phat trien manh, giai oan nay hau het cac mau hanchetaoprotocormmachchuyeutaochoi.Sosanhgia cac moi trng, th moi trng K5 tao them nhieu choi mi hn, cacchoimicomauxanhmt,vakhibatchoithchoixanh tot va ong loat hn hai moi trng con lai, co mot so choi co re nho, mau trang, dai t 0,1-0,3 cm. Co s khac biet ro ret ve so lng choi mi, chieu cao choi va chieu dai la gia cac moi trng ch co kch thch to BA, va cac moi trng co BA va IAA. cac moi trng ch co BA, sang en tuan th 4, cac protocorm mi bat choi, nhng so lng rat t, choi phat trien cham, va thap. Cac moi trng nay ch chu yeu tao them protocorm mi, t tao choi. Trong khi o, cac moi trng co BA va IAA, cac mau cay chu yeu tao choi, han che viec tao thanh protocorm. Nh vay, chung toi nhan thay, s ket hp gia BA va IAA th co li hn cho viec tao choi mi, phat trien cay con e s dung cho cac th nghiem ve sau. Tom lai:-oivithnghiemnhanprotocorm,thsdungmoitrngK2 vaK5eutot,nhngneusdungmoitrngK5,thsetao thanhnhieuchoimihn,cochieucao,vathchhpchoth nghiem ra re sau o. -NeusdungmoitrngK5choquatrnhnhangiongthheso nhan la: +~ .~+ :: ++- -: .~ +~ .~+ . .y-+ .. :+ ~+ y-+ ~+ 40 Cum choi ban au c tach lam 3, cay chuyen sang moi trngK5.Sau4tuan,tmotcumchoi,thuc4choicao 3-4cm.Nhvayvematlythuyet,tatnhchesonhan choi la 3x412 choi/nam. Sau 6 tuan,caccayconcochieu cao 3-4 cm va co 2-3 la c tach ra e a vaothnghiemtaore.Cacchoithapcungctachra,tieptucavaomoi trng K5, e phat trien chieu cao. Cac protocorm con lai cung c tach ra va a vao moi trng K2. +~ .~+ :: ++- -: .~ +~ .~+ . .y-+ .. :+ ~+ y-+ ~+ 41 +~ .~+ :: ++- -: .~ +~ .~+ . .y-+ .. :+ ~+ y-+ ~+ 42 V.Th nghiem 5: Nhng cay con trong th nghiem 4 co chieu cao t 3-4 cm, co t 2-3 la c cay vao moi trng ra re, sau 4 tuan theo doi ket qua ghi nhan c nh sau: Kch thch to Nong o (ppm) Ky hieuSo lng reChieu dai re (cm) Ty le cay ra re(%) Ghi chu 0K01-30,3-0,763,9Phat trien chieu cao. 0,1KN12-40,7-283,33Cay thap, phat trien khong eu. 0,5KN24-62-494,44La dai, xanh, phat trien tot va ong eu. NAA 1KN33-41-590,22Than om, la va re dai. 0,1KI11-30,5-1,262,5t re, la t va ngan.0,5KI22-31-4,569,44Phat trien tot,nhng khong eu, than thap, re dai. IAA 1KI31-31,5-3,566,67Cay thap, re t, la dai. 0,1KB12-42-377,78t la, phat trien khong eu. 0,5KB21-31-2,591,67La mau xanh phat trien tot, ong eu. IBA 1KB31-31-572,22Nhieu la, than ngan, re dai. Bang 6: Ket qua th nghiem tao re tren moi trng KnudsonC, bo sung NAA, IAA, IBA vi cac nong o khac nhau (sau 4 tuan). -Haituansaukhicayvaomoitrngrare,caybatautang chieu cao va re xuat hien. -Sau4tuan,rephattrienmanh,nhngtuytheotngloaimoi trng ma re, la va phat trien khac nhau. +~ .~+ :: ++- -: .~ +~ .~+ . .y-+ .. :+ ~+ y-+ ~+ 43 Da vao bang ket qua chung toi nhan thay: -Trong moi trng oi chng K0 , sau 4 tuan, choi ra re cham, ch co 1-3 re va chieu dai t 0,3-0,7 cm. Trong moi trng co bo sung kch thch to, kch thc cay va re co s bien oi ro ret tuy theo tuy loai kch thch to va nong o. -Trong moi trng bo sung kch thch to IAA, hau nh khong co s chenh lech ty le cay ra re, ty le thap ch tren 60% . So lng re khoang 1-3, nhng map va dai. Cay con thap, so lng la t, 2-3 la, la co mau xanh am. -Cac moi trng KB1, KB2, KB3, ty le cay ra re kha cao, va co s chenh lech ro rang. Moi trng KB2 co ty le cay ra re cao hn, nhng re ngan hn va co t la hn so vi moi trng KB3.-TrongmoitrngKN1,KN2,KN3,cotylecayrarecao.Cac cayconconhieula,ladai,comauxanhmt.Cayconcao, khoe co nhieu re, va dai. Trong moi trng KN2, ty le cay ra re cao nhat (94,44%), re dai, map, co mau xanh. Cay con co nhieu la,khoemanhsephattrienrattotkhichuyensanggiaioan chuyen tiep trc khi ra vn m. Nhnchung,ditacdungcuacackchthchtoNAA,IAA,IBAla cac auxin, se gay ra s giam o pH trong thanh te bao nen hoat hoa cac enzym phan huy cac polysaccharide la pectinmetylesteraza lien ket gia cac si cellulose lam chung long leo. Vach te bao mem ra, trnenlongleohnlamtebaokeodaira.Doovihamlng auxintangdanthsolngvachieudairecungtangtheo.Trong cacauxinthNAA,IBAcsdungnhieuhntrongmoitrng nuoicaymo.IAAtcsdungvtbenkhihapkhtrungmoi trng [10]. Tom lai moi trng totnhat cho s tao re lan Dendrobium la moi trng KnudsonCbosung0,5ppmNAA.Trongmoitrngnaycaycothegia tang chieu cao, so lng re va cng cap hn. sau khi cay tao c bo re cngcap,caykhoemanhthcotheacayvaogiaioanchuyentiep trc khi em ra trong ngoai vn. +~ .~+ :: ++- -: .~ +~ .~+ . .y-+ .. :+ ~+ y-+ ~+ 44 Hnh 4.5: Cay ra re tren moi trng KnudsonC vi cac nong o va kch to khac nhau, sau 4 tuan. +~ .~+ :: ++- -: .~+ - ++ . .y-+ .. :+ ~+ y-+ ~+36 PHAN V: KET LUAN E NGH I.Ket luan T nhng ket qua at c, chung toi a ra mot so ket luan sau: Mau kh trung la qua lan Dendrobium con xanh. V mauhat gieo nam ben trong, nen neu tien hanh ra mau ky va ot bang con 1 lan th co the loai bo hoan toan cac tac nhan gay nhiem. Hat khong b anh hng va khi gieo vao moi trng se phuc hoi rat nhanh. Moi trng MS co 1 ppm BA cho s nay mam cua hat lan Dendrobium tot hn so vi moi trng KnudsonC. S tao protocorm trong moi trng KnudsonC co bo sung kch thch to BA la tot nhat.oi vi th nghiem nhan protocorm s dung moi trng KnudsonC bo sung 2ppmBAva0,5ppmIAA,thsetaothanhnhieuchoimihn,cochieu cao, va thch hp cho th nghiem ra re sau o. Vi he so nhan choi la 3x412 choi/nam. Moi trng tot nhat cho s tao re lan Dendrobium la moi trng KnudsonC bo sung 0,5ppm NAA. T ket qua nghien cu tren chung toi a ra quy trnh nhan giong lan Dendrobium bang phng phap gieo hat in-vitro nh sau: +~ .~+ :: ++- -: .~+ - ++ . .y-+ .. :+ ~+ y-+ ~+37 Hnh 5.1: Quy trnh nhan giong lan Dendrobium bang phng phap gieo hat in-vitro. +~ .~+ :: ++- -: .~+ - ++ . .y-+ .. :+ ~+ y-+ ~+38 II.e ngh: Do thi gian thc hien e tai co han nen chung toi ch mi hoan thanh c giai oan gieo hat va nhan giong in-vitro. Trong thi gian ti, neu co ieu kien chung toi se tien hanh giai oan a cay con ra vn m va tiep tuc nghien cu cac th nghiem khac nh: -Khao sat anh hng o chn cua qua en s nay mam cua hat. -So sanh kha nang nhan choi mi so vi cac phng phap nhan giong lan Dendrobium khac. -Ket hp vi ky thuat lai giong hu tnhnham a ra nhng giong lan mi.