I-Science Behind Tissue Culture Plants: I-Micropropagation ne-Cloning Techniques

Isingeniso:

Izitshalo ze-tissue culture ziye zashintsha umhlaba we-horticulture nezolimo. Banikeza izinzuzo eziningi, okuhlanganisa ukukhiqizwa kwezitshalo ezingenazo izifo kanye nezakhi zofuzo ngezinga elikhulu. Le ndlela, eyaziwa ngokuthi i-micropropagation, ivumela ukuphindaphindeka ngokushesha nangempumelelo kwezitshalo, kuthuthukise isivuno sezitshalo futhi kusheshiswe inqubo yokongiwa kwezinhlobo zezitshalo. Kulesi sihloko, sizocubungula isayensi ngemuva kwezitshalo ze-tissue culture, sihlole indlela ye-micropropagation, futhi sembule amasu okuhlanganisa ahlakaniphile asetshenziswa kule nqubo. Ngokuqonda lezi zindlela, singabonga umthelela omkhulu izitshalo zesiko lezicubu ezinawo ezimbonini ezahlukahlukene namandla azo entuthuko yesikhathi esizayo.

Ukuqonda Isiko Lwezicubu

Isiko lezicubu, okubizwa nangokuthi i-in vitro propagation noma micropropagation, ibandakanya ukukhula kwamaseli ezitshalo noma izicubu endaweni elawulwayo ngaphandle kwendawo yazo yemvelo. Isebenza njengethuluzi elinamandla lokusabalalisa nokwandisa izitshalo okunzima noma eziphuza ukuzala ngezindlela zendabuko, njengembewu noma ukusikwa. Emasikweni ezicubu, izingcezu ezincane zezicubu zezitshalo, njengamaqabunga noma izingxenye zesiqu, zikhuliswa endaweni enomsoco ngaphansi kwezimo eziyinyumba. Lezi zicubu zikhethwa ngokucophelela, kuqinisekiswe ukuthi zinezici ezifiselekayo, njengokumelana nezifo noma isivuno esikhulu, ukuze zisakazeke.

Inqubo iqala ngokuvala inzalo i-explant, izicubu ezizokhuliswa, kusetshenziswa inhlanganisela yezinto ezibulala amagciwane. Lesi sinyathelo sibalulekile ukuze kuqedwe noma yimaphi amagciwane angase angcolise, angakhinyabeza ukukhula kwamasiko ezicubu. Uma sekufakwe inzalo, isitshalo sifakwa endaweni yesiko equkethe izakhamzimba ezibalulekile, amahomoni, nezilawuli zokukhula, ezikhuthaza ukuhlukana kwamaseli, ukwakheka kwamahlumela, nokuthuthukiswa kwezimpande.

Indawo esetshenziselwa isiko lamathishu ngokuvamile iqukethe ingxube kasawoti e-basal, ehlanganiswe namavithamini, ama-amino acid, noshukela. Amahomoni afana nama-auxin nama-cytokinin abalulekile ekulawuleni ukukhula nokuhlukaniswa kwezicubu zezitshalo. Inhlanganisela elinganiselayo ngokucophelela yala mahomoni ibalulekile ukuze kushukumise ukwakheka kwamahlumela nezimpande ezintsha, okuvumela ukukhiqizwa kwezitshalo eziphelele, ezinempilo.

Inqubo ye-Micropropagation

I-Micropropagation, ingxenye ebalulekile yesiko lezicubu, yenza ukuphindaphindeka ngokushesha kwama-plantlets. Le nqubo ihilela izinyathelo ezimbalwa ezihlukene, ezihlanganisa ukuqaliswa, ukuphindaphinda, ukuqothula, kanye nokulinganisa.

Ukuqaliswa: Isigaba sokuqala sibonisa ukusungulwa kwamasiko ase-aseptic. Izitshalo ezikhulayo zikhethwa ngokucophelela ezitshalweni ezinempilo, ezingenazo izifo futhi zifafazwe amagciwane ukuze kuqedwe ukungcola okungenzeka. Ama-explants abe esebekwa endaweni ye-culture ukuze aqalise ukukhula kwama-callus amancane, angahlukanisiwe.

Ukuphindaphinda: Uma isigaba sokuqala sesiphumelele, isinyathelo esilandelayo ukuphindaphinda. Phakathi nalesi sigaba, ama-callus mass afakwa endaweni entsha ukuze akhuthaze ukukhula kwamahlumela amaningi. Lawa amahlumela abe esehlukaniswa bese edluliselwa ngawodwa endaweni entsha yesiko, okuvumela ukuphindaphindeka okwengeziwe kanye nokwakheka kwezithombo.

Ukuqhakaza: Ngemva kokuthola ukuphindaphindeka kwehlumela okuphawulekayo, izitshalo zezitshalo zidluliselwa endaweni yokuhluma. Le medium iqukethe ukugcwala okuphezulu kwama-auxins, okukhuthaza ukukhula kwezimpande. Izitshalo zigadwa ngokucophelela ukuze kuqinisekiswe ukukhula okufanele kwezimpande kanye nokumiswa ngaphambi kokuba zidluliselwe emhlabathini noma endaweni yokukhula efanele.

I-Acclimatization: Isinyathelo sokugcina se-micropropagation ukujwayela. Ama-plantlets, manje anezimpande ezithuthuke kahle, kancane kancane avezwa endaweni yangaphandle. Ekuqaleni, zifakwa endaweni elawulwayo, njenge-greenhouse, ukuze kugwenywe ukushintsha okungazelelwe kwezinga lokushisa nomswakama. Ngokuhamba kwesikhathi, ama-plantlets ajwayela indawo yawo entsha, agcine esevuthiwe futhi akwazi ukukhula ngokuzimela.

Amasu Okuhlanganisa Ezitshalweni ZeSiko Lwezicubu

I-Cloning ngokusebenzisa isiko lezicubu kunikeza indlela ephumelelayo nethembekile yokukhiqiza izitshalo ezinezici ezifiselekayo. Kusetshenziswa izindlela ezahlukahlukene zokuhlanganisa, kuye ngohlobo lwesitshalo kanye nemiphumela efiselekayo.

I-Cloning Yezitshalo Ezivuthiwe: Le nqubo ihilela ukusetshenziswa kwezicubu zezitshalo ezivuthiwe, njengamaqabunga noma iziqu, ukuqalisa amasiko ezicubu. Ama-explants athathwa esitshalweni esikhule ngokugcwele futhi abekwe endaweni yesiko, ekhuthaza ukwakheka kwamangqamuzana angahlukanisiwe angaqhubeka nokuvuselela ezitshalweni eziphelele. Le ndlela inenzuzo lapho usakaza izinhlobo ezithatha isikhathi eside ukufinyelela ukuvuthwa kokuzala.

I-Meristem Culture: Isiko le-Meristem ligxile ekutshalweni kwama-apical meristems—isifunda esisekupheleni kwehlumela elikhulayo elinamaseli anamandla akhethekile okukhula. Lawa maseli anamandla okukhiqiza kabusha izitshalo eziphelele. Ngokuhlakulela ama-meristems, kungenzeka ukuthola izitshalo zeqiniso-to-type, ezifana ngokwemvelo nesitshalo esingumama, okuqinisekisa ukulondolozwa kwezici ezithile.

Isiko Lombungu: Isiko lombungu libandakanya ukunqunywa noma ukukhishwa kwemibungu engavuthiwe embewini, okulandelwa ukubekwa kwayo endaweni efanele yesiko. Le nqubo iwusizo ikakhulukazi ekusakazeni izitshalo ezikwazi ukukhula kwembewu ephansi noma lezo ezibonisa ukulala kwembewu. Iqinisekisa ukuthi izici ezibalulekile noma eziyingqayizivele ezikhona emibungu zigcinwa ngempumelelo futhi zande.

I-Callus Culture: I-Callus Culture iyindlela esebenzisa ikhono eliyingqayizivele lamaseli angahlukanisiwe ukuze enze i-callus-inqwaba yamaseli ahlukanisayo. Izitshalo ezithathwe ezingxenyeni ezihlukahlukene zesitshalo zifakwa endaweni yesiko, zikhuthaze ukukhula kwe-callus. I-Callus Culture iyindlela ephumelelayo yokusabalalisa izinhlobo ezingenazo izakhiwo ezikhethekile zokuzala, futhi ivumela ukukhiqizwa kwenani elikhulu lezitshalo.

I-Somatic Embryogenesis: I-Somatic embryogenesis ibandakanya ukwakheka kwemibungu evela kumaseli e-somatic, okungamaseli angazali esitshalo. Inqubo ye-somatic embryogenesis iqala ngokuhlakulela amaseli ngendlela efanelekile, evumela ukukhula kombungu. Le nqubo inamandla amakhulu njengoba ivumela ukukhiqizwa kwenani elikhulu lemibungu, ngayinye engase iveze isitshalo esifana nofuzo.