Ibikoresho by'ibimera n'udukoko twanduza indwara
Ihuriro ry’amashaza rihuza ubwoko bw’amashaza rizwi nka sorghum conversion population (SCP) ryatanzwe na Dr. Pat Brown wo muri Kaminuza ya Illinois (ubu iri muri UC Davis). Ryavuzwe mbere kandi ni icyegeranyo cy’imirongo itandukanye yahinduwe ihinduka idafite ubushobozi bwo kumenya igihe cy’izuba n’uburebure buto kugira ngo byorohereze imikurire n’iterambere ry’ibimera mu bidukikije bya Amerika. Imirongo 510 yavuye muri ubu bwoko yakoreshejwe muri ubu bushakashatsi nubwo bitewe n’imikurire mibi n’ibindi bibazo byo kugenzura ubuziranenge, si imirongo yose yakoreshejwe mu gusesengura imiterere yose uko ari itatu. Amaherezo amakuru yavuye mu mirongo 345 yakoreshejwe mu gusesengura igisubizo cya chitin, imirongo 472 mu gisubizo cya flg22, na 456 mu kurwanya TLS.B. cookeiUbwoko bwa LSLP18 bwakuwe kuri Dr. Burt Bluhm wo muri Kaminuza ya Arkansas.
Igipimo cy'igisubizo cya MAMP
Muri ubu bushakashatsi hakoreshejwe MAMP ebyiri zitandukanye, (Genscript catalog# RP19986), na chitin. Ibimera by'amasaka byatewe mu bice byashyizwe ku butaka bwuzuyemo ubutaka (33% Sunshine Redi-Earth Pro Growing Mix) mu busitani. Ibimera byavomerewe umunsi umwe mbere y'ikusanyirizo ry'ibihingwa kugira ngo hirindwe ubushuhe bwinshi ku munsi wo gukusanya.
Imirongo yakozwe ku buryo butaziguye, kandi kubera impamvu z’ibijyanye n’ikoranabuhanga, yatewe mu byiciro by’imirongo 60. Kuri buri murongo, hatewe "inkono" eshatu zifite imbuto ebyiri kuri buri murongo. Ibice byakurikiyeho byatewe vuba cyane nyuma yuko itsinda ryabanje rimaze gutunganywa kugeza igihe abaturage bose bapimwe. Hakozwe igerageza kabiri kuri MAMP zombi, aho genotypes zombi zasubiwemo ku buryo butaziguye muri buri cyiciro muri ibyo byiciro bibiri.
Isuzuma rya ROS ryakozwe nk'uko byasobanuwe mbere. Muri make, kuri buri murongo, imbuto esheshatu zatewe mu nkono 3 zitandukanye. Mu mbuto zavuyemo, eshatu zatoranijwe hashingiwe ku buryo bumwe. Imbuto zisa n'izidasanzwe cyangwa ndende cyane cyangwa ngufi kuruta nyinshi ntizakoreshejwe. Uduce duto tw'amababi tune dufite umurambararo wa mm 3 twakuwe mu gice kinini cy'ikibabi cya 4 cy'ibimera bitatu bitandukanye by'amasaka by'iminsi 15. Uduce duto kuri buri kibabi cy'ibimera bibiri n'uduce duto tubiri ku kimera kimwe, aho utwa kabiri tuba ari two dugenzura amazi (reba hepfo). Uduce duto twashyizwe ku mazi ku gipimo cya 50 µl H20 mu isahani y'umukara ifite amariba 96, dufunze n'agapfundikizo ka aluminiyumu kugira ngo wirinde koherezwa n'urumuri, kandi tubikwa ku bushyuhe bw'icyumba ijoro ryose. Mu gitondo cyakurikiyeho, umuti wo gusubiza wakozwe hakoreshejwe 2 mg/ml ya chemiluminescent probe L-012 (Wako, catalog # 120-04891), 2 mg/ml ya horseradish peroxidase (Ubwoko bwa VI-A, Sigma-Aldrich, catalog # P6782), na 100 mg/ml ya Chitin cyangwa 2 μM ya Flg22. 50 µl by'uyu muti wo gusubiza wongewe ku mariba atatu muri ane. Iriba rya kane ryari uburyo bwo kugenzura, aho hongewemo umuti wo gusubiza utarimo MAMP. Hari kandi amariba ane y'ubusa arimo amazi gusa muri buri sahani.
Nyuma yo kongeramo igisubizo cy’ibizamini, urumuri rwapimwe hakoreshejwe SynergyTM 2 multi-detection microplate reader (BioTek) buri munota 2 mu gihe cy'isaha 1. Isoma ry'amapine ripima urumuri buri munota 2 muri iki gihe cy'isaha 1. Igiteranyo cy'ibipimo 31 byose cyabazwe kugira ngo haboneke agaciro ka buri riba. Agaciro kagereranyijwe k'igisubizo cya MAMP kuri buri bwoko bw'ibinyabuzima kabazwe nka (ikigereranyo cy'agaciro k'urumuri rw'amapine atatu y'igerageza—agaciro k'amapine y'igerageza) - gukuramo agaciro k'ikigereranyo k'amapine y'igerageza. Agaciro k'amapine y'igerageza kari hafi ya zeru.
Uduce tw'amababi yaNicotiana benthamiana, umurongo umwe w’amasaka usubiza neza (SC0003), n’umurongo umwe w’amasaka usubiza neza (PI 6069) nabyo byashyizwe mu buryo bwo kugenzura muri buri sahani y’amasōko 96 kugira ngo bigenzurwe ubuziranenge.
B. cookeigutegura no gutera urushinge
B. cookeiInkongoro yateguwe nk'uko byasobanuwe mbere. Muri make, imbuto z'amasaka zashyizwe mu mazi iminsi itatu, zozwa, zishyirwa mu macupa ya litiro 1 y'udupira tw'ubwoko bwa conical hanyuma zigashyirwa mu gihe cy'isaha imwe kuri 15psi na 121 °C. Hanyuma izo nkoko zateweho mililitiro zigera kuri 5 za mycelium ya macerated yavuye mu miterere mishya yaB. cookeiLSLP18 irakura ikarekera mu byumweru 2 ku bushyuhe bw'icyumba, ikazunguza amacupa buri minsi 3. Nyuma y'ibyumweru 2, imbuto z'amasaka zanduye zarumikwaga mu kirere hanyuma zikabikwa kuri dogere selisiyusi 4 kugeza igihe zitewe mu murima. Urushinge rumwe rwakoreshejwe mu igerageza ryose kandi rukorwa bushya buri mwaka. Mu gutera, imbuto 6-10 zanduye zashyizwe mu gishishwa cy'ibimera by'amasaka bifite ibyumweru 4-5. Udukoko twaturutse kuri izi bishingwe twateye ubwandu mu bimera by'amasaka bikiri bito mu cyumweru kimwe.
Gutegura imbuto
Mbere yo gutera mu murima imbuto z'amasaka zaterwaga umuti urwanya udukoko, udukoko twica udukoko, n'umuti urinda indwara urimo ~ 1% umuti urwanya indwara wa Spirato 480 FS, umuti urwanya indwara wa Sebring 480 FS, umuti urinda indwara wa Sorpro 940 ES 3%. Hanyuma imbuto zumishwa mu kirere mu gihe cy'iminsi 3, ibyo bikaba byaratumye uru ruvange rutwikira imbuto. Umuti urinda indwara wemeye gukoresha umuti wa Dual Magnum nk'umuti mbere yo kumera.
Isuzuma ry'uko imiterere y'amabara arwanya imiterere y'amabara ku ibara rya Target Leaf ihagaze
SCP yatewe muri sitasiyo y’ubushakashatsi ku bihingwa i Clayton, muri leta ya NC ku ya 14 kugeza 15 Kamena 2017 na 20 Kamena 2018 mu buryo butunguranye, hakozwe imiterere ibiri y’igerageza muri buri kibazo. Igerageza ryatewe mu mirongo imwe ya metero 1.8 ifite ubugari bwa metero 0.9 hakoreshejwe imbuto 10 kuri buri gice cy’ubutaka. Imirongo ibiri y’imbibi yatewe hirya no hino ku nkengero za buri gice cy’igerageza kugira ngo hirindwe ingaruka mbi. Igerageza ryatewe ku ya 20 Nyakanga 2017 na 20 Nyakanga 2018, aho ibimera by’amasaka byari ku rwego rwa 3 rwo gukura. Amanota yafashwe ku gipimo cya rimwe kugeza ku icyenda, aho ibimera bitagaragazaga ibimenyetso by’indwara byahawe amanota nk’ibimera icyenda kandi byapfuye burundu. Amanota abiri yafashwe mu 2017 n’amanota ane mu 2018 atangira ibyumweru bibiri nyuma yo guterwa buri mwaka. sAUDPC (agace gasanzwe kari munsi y’umurongo w’izamuka ry’indwara) yabazwe nk’uko byasobanuwe mbere.
Igihe cyo kohereza: Mata-01-2021