Iyini i-Inoculation ku-Plant Tissue Culture?

Isingeniso:

Isiko lezicubu zezitshalo kuyindlela ebalulekile esetshenziswa ekusakazeni nasekutshalweni kwezitshalo endaweni elawulwayo. Kubandakanya ukukhula kwamaseli ezitshalo, izicubu, noma izitho ezingaphansi kwezimo eziyinyumba endaweni enomsoco. Ukujova kuyisinyathelo esibalulekile empumelelweni yezicubu zezitshalo, esethula isitshalo endaweni eyinyumba yesiko. Lesi sihloko singena emcabangweni wokujova esikweni lezicubu zezitshalo, sihlola inhloso, izindlela, kanye nokubaluleka kwawo emkhakheni wokusabalalisa izitshalo.

Ukuqonda I-inoculation ku-Plant Tissue Culture:

I-inoculation isho inqubo yokwethula izinto zezitshalo, njengezimila noma amasiko e-callus, endaweni eyinyumba yesiko. Lesi sinyathelo sivumela impahla yesitshalo ukuthi izimise endaweni elawulwayo, ivumele ukukhula nokuphindaphindeka kwayo. I-inoculation ngokuvamile yenziwa ku-laminar flow hood, ihlinzeka ngendawo yokusebenza eyinyumba ukuvimbela ukungcola.

Inhloso ye-Inoculation:

Inhloso eyinhloko yokujova kusiko lwezicubu zezitshalo ukuqala ukukhula kwezitshalo endaweni elawulwayo. I-culture medium oyinyumba inikeza zonke izakhi ezidingekayo, amahomoni, nezilawuli zokukhula ezibalulekile ekukhuleni kwezitshalo. Ngokwethula izinto zezitshalo kule ndawo, inqubo yesiko lezicubu ingaqalwa ngempumelelo.

I-inoculation iphinde isebenze nezinye izinjongo ezibalulekile. Ivumela ukuphindaphindeka kwezitshalo ngokukhiqizwa kwamahlumela amaningi, izimpande, noma imibungu. Isiza ekukhiqizeni isitshalo esingenazifo ngokuqeda amagciwane kanye nezinambuzane. I-inoculation iphinde isetshenziselwe ukuguqulwa kwezakhi zofuzo, lapho izakhi zofuzo zangaphandle zifakwa ezicutshini zezitshalo, okuholela ekuthuthukisweni kwezitshalo eziguqulwe izakhi zofuzo.

Izindlela ze-inoculation:

Kunezindlela eziningana zokujova ezisetshenziselwa isiko lezicubu zezitshalo, kuye ngokuthi hlobo luni lwezitshalo kanye nomphumela oyifunayo. Nazi izindlela ezimbalwa ezivame ukusetshenziswa:

1. Ukwenziwa inzalo ebusweni:

Ukuvala inzalo ebusweni kuyisinyathelo esibalulekile ngaphambi kokugonywa, ikakhulukazi lapho kusetshenziswa izitsha ezivela endaweni yemvelo. Kuhilela ukususwa kwezinto ezingcolile, njengesikhunta, amabhaktheriya, namanye amagciwane, endaweni yesitshalo. Indlela ejwayeleke kakhulu yokuvala inzalo ebusweni ukusetshenziswa kwamakhemikhali, njenge-sodium hypochlorite noma i-ethanol. Ama-explants acwiliswe kusixazululo sokubulala amagciwane isikhathi esithile, okuqinisekisa ukuqedwa ngokuphelele kokungcola ngaphandle kokulimaza isitshalo.

2. I-Meristem Culture:

Isiko le-Meristem lihilela ukujova kwe-apical meristem, okuyizicubu ezihlukanisayo ngokuqhubekayo ekugcineni kwamahlumela noma izimpande. Izicubu ze-Meristematic zinamandla amakhulu okukhula futhi zivame ukusetshenziselwa ukunqoba izinkinga zokungcola. Ngokujova ama-meristems, ukukhula kwezicubu ezingangcolisiwe kuyaqinisekiswa, okuholela ekukhiqizeni izitshalo ezingenazifo. Isiko le-Meristem liwusizo ikakhulukazi ekusakazeni izinhlobo zezitshalo ezibalulekile noma ezingavamile, lapho ukulondoloza ubuqotho bazo bofuzo kubalulekile.

3. I-Callus Culture:

Isiko le-Callus lihilela ukufakwa kwezicubu ze-callus, ezingahlukanisiwe, amangqamuzana akhula ngobuningi. Ngale ndlela, izitsha ezincane noma izingxenye zezitshalo zifakwa endaweni yesiko elicebile izakhi eziqukethe izilawuli zokukhula kwezitshalo. Ngokuhamba kwesikhathi, amaseli e-explant ayanda futhi enze i-callus, engase ifakwe ngaphansi kwe-subcultured ukukhiqiza izitshalo eziningi. Isiko se-Callus sisetshenziswa kabanzi ekusakazeni izinhlobo zezitshalo ezifanayo zofuzo, njengoba kuvumela ukukhiqizwa kwenani elikhulu lezitshalo kusuka kumthamo omncane wezinto zokuqala.

4. Isiko Lombungu:

Isiko le-embryo livame ukusetshenziselwa ukusabalalisa izitshalo ezinembewu enemibungu engathuthukisiwe noma imbewu enemibungu elele kakhulu. Ngale ndlela, imibungu ihlukaniswa nembewu futhi ibekwe endaweni yesiko eliqukethe izilawuli ezithile zokukhula. Le nqubo ivumela imibungu ukuthi iqhume futhi ikhule ibe izitshalo zezitshalo ngaphandle kwembewu, iqinisekise ukukhula kwayo ngempumelelo kanye nokuzala. Isiko le-embryo libalulekile ekusakazeni kwezinhlobo ezithile zezitshalo, obekungaba inselele ukusabalala ngezindlela zendabuko.

5. Isiko Lomzimba:

Inqubo yesitho somzimba ihlanganisa ukujova kwazo zonke izitho zezitshalo, njengezimpande, amahlumela, noma amaqabunga, endaweni yesiko. Le ndlela ivumela ukukhula nokuvuselelwa kwazo zonke izitshalo ezivela ezingxenyeni zesitshalo ngasinye. Isitho somzimba siwusizo ikakhulukazi ekukhiqizeni ama-plantlets kusuka ezingxenyeni zezitshalo, njengokusikwa kwamaqabunga noma izingxenye zezimpande. Ngokuhlinzeka ngezakhi ezidingekayo kanye nezilawuli zokukhula, isiko lesitho sisiza ukukhula nokuthuthukiswa kwezicubu zezitshalo zibe izitshalo eziphelele.

Ukubaluleka Kokufakwa Kwama-Inoculation Kusiko Lezicubu Zezitshalo:

I-inoculation idlala indima ebalulekile empumelelweni yesiko lezicubu zezitshalo. Ngaphandle kwamasu afanele wokujova, isitshalo singase singakhuli, okuholela ekungcoleni noma ekufeni kwezicubu. Ngokwethula isitshalo endaweni eyinyumba yesiko, amathuba okungcola ancishiswa kakhulu, okuqinisekisa ukukhula kwezicubu ezinempilo nezingangcolisiwe.

Ngaphezu kwalokho, ukujova kuvumela ukusabalala kwezitshalo ngezinga elikhulu. Ngokusebenzisa amasu afanele we-subculturing, isimila esisodwa singaveza izimila eziningi. Le ndlela yokusabalalisa ngobuningi iwusizo ikakhulukazi ezikhungweni zezentengiselwano nabalimi bezitshalo, okubenza bakwazi ukukhiqiza izitshalo eziningi ngesikhathi esifushane.

I-inoculation ibalulekile futhi ekusetshenzisweni kwezindlela zobunjiniyela bezakhi zofuzo kusiko lezicubu zezitshalo. Ngokufaka izakhi zofuzo zangaphandle ezicutshini zezitshalo ngokuzijova, ososayensi bangakha izitshalo ezinezici ezifiselekayo, njengokumelana nezinambuzane, izifo, noma isivuno esithuthukisiwe. Lezi zitshalo ezishintshwe izakhi zofuzo zinemithelela ebalulekile kwezolimo, ezinomthelela ekukhiqizeni izitshalo ezithuthukisiwe kanye nokusimama.

Sengiphetha, ukujova kuyisinyathelo esibalulekile esimisweni sezicubu zezitshalo esivumela ukukhula, ukuphindaphindeka, kanye nokuguqulwa kofuzo kwezitshalo endaweni elawulwayo. Izindlela ezahlukahlukene zokujova, njengokuvala inzalo ebusweni, isiko le-meristem, isiko le-callus, isiko le-embryo, kanye nesiko lesitho, zinikeza izinketho zokusatshalaliswa kwezinhlobonhlobo zezitshalo zezitshalo. Ngokuqonda nokusebenzisa izindlela ezifanele zokujova, ososayensi nabalimi bezitshalo bangafinyelela isiko eliphumelelayo lezicubu zezitshalo, libe nesandla ekuthuthukisweni kwezitshalo ezingenazo izifo, ukusakazeka ngobuningi, kanye nentuthuko yobunjiniyela bezakhi zofuzo.