Inyoni ya pinusi ni inyamaswa zimuka mu kato zizwiho gutera igihombo gikomeye mu bukungu mu miterere y’ibinyabuzima by’ishyamba rya pinusi. Ubu bushakashatsi busuzuma imikorere y’inyoni za halogenated zirwanya inyoni za pinusi n’uburyo zikora. Ibikorwa byo kwica inyoni bya 5-iodoindole na avermectin (gugenzura neza) barwanya inyoni za pinusi byari bisa kandi biri hejuru ku gipimo gito (10 μg / mL). 5-iodoindole yagabanyije ububobere, imikorere yo kubyara, impfu z’urusoro n’iz’inkoko, hamwe n’imyitwarire y’uturemangingo. Uburyo ligande zikorana n’udukoko tw’inkoko dufite glutamate-gated chloride channel receptors zishyigikira igitekerezo cy’uko 5-iodoindole, kimwe na avermectin, yihambira neza ku gice cy’inyuni zikora. 5-Iodoindole yanateye impinduka zitandukanye mu miterere y’inyuni, harimo no kugwa/gucika kw’ingingo kudasanzwe no kwiyongera kwa vacuolization. Ibi byavuye mu bushakashatsi bigaragaza ko vacuole zishobora kugira uruhare mu rupfu ruterwa na methylation ya nematode. Icy'ingenzi ni uko 5-iodoindole itari ifite uburozi ku bimera byombi (amashu n'amaradishe). Bityo, ubu bushakashatsi bwerekana ko gukoresha iyoodoindole mu bihe by'ibidukikije bishobora kurwanya kwangirika kw'ibiti bya pinusi.
Indwara y’ibiti bya pinusi (Bursaphelenchus xylophilus) iri mu bwoko bwa nematode zo mu giti cya pinusi (PWN), zikunda kwimuka zizwiho kwangiza cyane ibidukikije ku rusobe rw’ibinyabuzima byo mu ishyamba rya pinusi1. Indwara ya pinusi (PWD) iterwa na nematode zo mu giti cya pinusi irimo kuba ikibazo gikomeye ku migabane myinshi, harimo Aziya n’Uburayi, kandi muri Amerika ya Ruguru, nematode yica ubwoko bwa pinusi bwazanywe1,2. Kugabanuka kw’ibiti bya pinusi ni ikibazo gikomeye mu bukungu, kandi icyizere cyo gukwirakwira kwabyo ku isi kirateye impungenge3. Ubwoko bukurikira bwa pinusi bukunze kwibasirwa cyane na nematode: Pinus densiflora, Pinus sylvestris, Pinus thunbergii, Pinus koraiensis, Pinus thunbergii, Pinus thunbergii, na Pinus radiata4. Nematode yo mu giti cya pinusi ni indwara ikomeye ishobora kwica ibiti bya pinusi mu byumweru cyangwa amezi nyuma yo kwandura. Byongeye kandi, icyorezo cya nematode zo mu giti cya pinusi gikunze kugaragara mu rusobe rw’ibinyabuzima bitandukanye, bityo hamaze gushyirwaho uruhererekane rw’indwara zidashira1.
Bursaphelenchus xylophilus ni nematode yo mu bwoko bwa "quarantine" ikomoka ku bimera byo mu muryango wa Aphelenchoidea na clade 102.5. Nematode irya ibihumyo kandi ikororoka mu bice by'ibiti by'ibiti bya pinusi, ikura mu byiciro bine bitandukanye by'ibihumyo: L1, L2, L3, L4 n'umuntu mukuru1,6. Mu gihe cy'ibura ry'ibiribwa, nematode yo mu bwoko bwa pinusi inyura mu cyiciro cyihariye cy'ibihumyo - dauer, itera indwara - agakoko ka pinusi (Monochamus alternatus) maze ijyanwa mu biti by'ibiti bya pinusi bizima. Mu binyabuzima bizima, nematode zihita zinyura mu bice by'ibimera zikarya uturemangingo tw'ibimera, ibi bigatera ingaruka nyinshi zo kwangirika, gushwanyagurika no gupfa mu mwaka umwe nyuma yo kwandura1,7,8.
Kurwanya udukoko twa pinusi tw’ibiti bya pinusi byamaze igihe kirekire ari ikibazo gikomeye, aho ingamba zo gushyira mu kato zatangiye mu kinyejana cya 20. Ingamba zigezweho zo kurwanya udukoko twa pinusi zirimo cyane cyane imiti ikoreshwa mu kuvura indwara, harimo no gutwika ibiti no gutera udukoko twa pinusi mu biti. Udukoko twa pinusi dukoreshwa cyane ni avermectin na avermectin benzoate, biri mu muryango wa avermectin. Iyi miti ihenze igira akamaro kanini mu kurwanya ubwoko bwinshi bwa pinusi kandi ifatwa nk'aho idafite ingaruka ku bidukikije9. Ariko, gukoresha iyi miti kenshi byitezwe ko bizatera igitutu cyo guhitamo ibintu bizatera kugaragara kwa pinusi twa pinusi, nk'uko byagaragaye ku dukoko twinshi dukunda udukoko, nka Leptinotarsa decemlineata, Plutella xylostella na pinusi za Trichostrongylus colubriformis na Ostertagia circumcincta, zagiye zitera imbere mu kurwanya avermectins10,11,12. Kubwibyo, imiterere yo kurwanya igomba kwigwa buri gihe kandi imiti ya pinusi igasuzumwa buri gihe kugira ngo haboneke ubundi buryo, buhendutse kandi butangiza ibidukikije bwo kurwanya PVD. Mu myaka ibarirwa muri za mirongo ishize, abanditsi benshi batanze igitekerezo cyo gukoresha ibikomoka ku bimera, amavuta y’ingenzi n’ibintu bihumanya nk’ibintu birwanya udukoko 13,14,15,16.
Duherutse kwerekana imikorere ya indole, molecule y’ibimenyetso hagati y’uturemangingo n’ibiri hagati y’uturemangingo, muri Caenorhabditis elegans 17. Indole ni ikimenyetso cyagutse mu turemangingo mu bidukikije bya mikorobe, igenzura imikorere myinshi igira ingaruka ku miterere ya mikorobe, ikorwa ry’uturemangingo, kudahinduka kwa plasmide, kudafata imiti, ikorwa rya biofilm, no kwandura 18, 19. Umurimo wa indole n’ibikomoka kuri yo ku zindi nematode zitera indwara ntabwo wizweho. Muri ubu bushakashatsi, twasesenguye imikorere ya nematode 34 zirwanya nematode za pinusi kandi dusobanura uburyo 5-iodoindole ikomeye cyane ikora hakoreshejwe mikorosikopi, ifoto ya time-lapse, n’igerageza rya molekile, kandi dusuzuma ingaruka z’uburozi bwayo ku bimera dukoresheje isuzuma ryo kumera kw’imbuto.
Mbere byavuzwe ko indole ifite ingano nini (>1.0 mM) igira ingaruka ku mikorobe17. Nyuma yo kuvura B. xylophilus (ibyiciro bitandukanye by'ubuzima) hakoreshejwe indole cyangwa 33 bitandukanye bya indole kuri 1 mM, urupfu rwa B. xylophilus rwapimwe hagaragazwa imikorobe mizima n'iyapfuye mu matsinda yagenzuwe n'ayavuwe. Indole eshanu zagaragaje imikorere ikomeye yo kwica imikorobe; kubaho kw'itsinda ryagenzuwe ryavuwe byari 95 ± 7% nyuma y'amasaha 24. Muri indole 34 zapimwe, 5-iodoindole na 4-fluoroindole kuri 1 mM byateje urupfu 100%, mu gihe 5,6-difluoroindigo, methylindole-7-carboxylate, na 7-iodoindole byateje urupfu rugera kuri 50% (Imbonerahamwe ya 1).
Ingaruka za 5-iodoindole ku miterere ya vacuole no ku mikorere ya nematode yo mu giti cya pinusi. (A) Ingaruka za avermectin na 5-iodoindole ku nematode z'abagabo bakuze, (B) amagi ya nematode yo mu cyiciro cya L1 na (C) imikoreshereze ya B. xylophilus, (i) vacuole ntizagaragaye mu isaha 0, ubuvuzi bwatumye (ii) vacuole, (iii) kwibumbira kwa vacuole nyinshi, (iv) kubyimba kwa vacuole, (v) kwibumbira kwa vacuole na (vi) kwibumbira kwa vacuole nini. Utwambi dutukura tugaragaza kubyimba kwa vacuole, utwambi tw'ubururu tugaragaza kwibumbira kwa vacuole n'utw'umukara tugaragaza vacuole nini. Uduce twa scale = 50 μm.
Byongeye kandi, ubu bushakashatsi bwanasobanuye inzira y’urupfu ruterwa na methane muri pinusi (Ishusho ya 4C). Urupfu rwa methane ni ubwoko bw’urupfu rw’uturemangingo rudakomoka ku ruhu rufitanye isano no kwibumbira kwa vacuole zigaragara za cytoplasmic27. Ubusembwa bw’imiterere bwagaragaye muri pinusi za pinusi busa n’aho bufitanye isano rya bugufi n’uburyo urupfu ruterwa na methane ruterwa. Isuzuma rya mikorosikopi mu bihe bitandukanye ryagaragaje ko vacuole nini zabayeho nyuma y’amasaha 20 zo guhura na 5-iodoindole (0.1 mM). Vacuole za mikorosikopi zagaragaye nyuma y’amasaha 8 zo kuvurwa, kandi umubare wazo wiyongereye nyuma y’amasaha 12. Vacuole nini nyinshi zagaragaye nyuma y’amasaha 14. Vacuole nyinshi zahujwe zagaragaye neza nyuma y’amasaha 12-16 zo kuvurwa, bigaragaza ko fusion ya vacuole ari yo shingiro ry’uburyo bwo gupfa bwa methane. Nyuma y’amasaha 20, vacuole nyinshi nini zabonetse mu nyo. Ibi byagaragaye ni raporo ya mbere ya metuosis muri C. elegans.
Mu nzoka zavuwe na 5-iodoindole, kwishyira hamwe no gucika kwa vacuole nabyo byagaragaye (Ishusho ya 5), nk'uko bigaragazwa no kunama kw'izo nzoka no kurekura vacuole mu bidukikije. Guhungabana kwa vacuole nabyo byagaragaye mu gice cy'igi cy'amagi, ubusanzwe L2 iguma ikora neza mu gihe cyo gusohora (Ishusho y'inyongera S2). Ibi biremeza uruhare rw'amazi yiyongera no kunanirwa kwa osmoregulation, ndetse no kwangirika kw'uturemangingo (RCI), mu gihe vacuole ikora kandi igacika (Ishusho ya 5).
Dutekereza ku ruhare rwa iyode mu miterere ya vacuole yagaragaye, twakoze ubushakashatsi ku mikorere ya sodium iyode (NaI) na iyode potasiyumu (KI) yo kwica udukoko. Ariko, ku gipimo cya (0.1, 0.5 cyangwa 1 mM), ntabwo byagize ingaruka ku kubaho kwa nematode cyangwa ku miterere ya vacuole (Ishusho y'inyongera S5), nubwo 1 mM KI yagize ingaruka nke ku miterere ya nematode. Ku rundi ruhande, 7-iodoindole (1 cyangwa 2 mM), kimwe na 5-iodoindole, yateje vacuole nyinshi n'imiterere y'imiterere (Ishusho y'inyongera S6). Iodoindole ebyiri zagaragaje imiterere isa ya phenotypic mu nematode za pine, mu gihe NaI na KI zitagaragaje. Igishimishije ni uko indole itateje ihinduka rya vacuole muri B. xylophilus ku gipimo cyapimwe (amakuru ntabwo yagaragajwe). Bityo, ibisubizo byemeje ko indole-iyode complex ari yo ituma B. xylophilus ikora vacuolization na metabolism.
Mu byapimwe ku bijyanye n’imikorere y’uturemangingo tw’indole, 5-iodoindole yari ifite igipimo cyo hasi cya -5.89 kcal/mol, ikurikirwa na 7-iodoindole (-4.48 kcal/mol), 4-fluoroindole (-4.33), na indole (-4.03) (Ishusho ya 6). Guhuza cyane kwa hydrogen kwa 5-iodoindole na leucine 218 bituma ikomeza gufatana kwayo, mu gihe ibindi bikomoka kuri indole byose bifatanya na serine 260 binyuze mu migozi ya hydrogen yo ku ruhande. Mu zindi iodoindole zakozwe mu buryo bwa modeli, 2-iodoindole ifite agaciro ko gufatana ka -5.248 kcal/mol, bitewe n’uko ifitanye isano nyamukuru na hydrogen na leucine 218. Izindi ngano zizwi zirimo 3-iodoindole (-4.3 kcal/mol), 4-iodoindole (-4.0 kcal/mol), na 6-fluoroindole (-2.6 kcal/mol) (Ishusho y’inyongera S8). Indole nyinshi za halogenated na indole ubwazo, usibye 5-iodoindole na 2-iodoindole, zigira isano na serine 260. Kuba hydrogen ifatanya na leucine 218 bigaragaza uburyo receptor-ligand ikorana neza, nk'uko byagaragaye kuri ivermectin (Ishusho y'inyongera S7), byemeza ko 5-iodoindole na 2-iodoindole, kimwe na ivermectin, bifatana neza n'aho receptor ya GluCL ikorera binyuze muri leucine 218 (Ishusho ya 6 n'Ishusho y'inyongera S8). Turasaba ko iyi mibonano ikenewe kugira ngo imiterere y'umwobo ufunguye wa GluCL ikomeze kandi ko mu gufatana neza n'aho receptor ya GluCL ikorera, 5-iodoindole, 2-iodoindole, avermectin na ivermectin bityo bigatuma umuyoboro wa ion ukomeza gufungura kandi bigatuma amazi yinjira.
Gushyira indole na halogenated indole kuri GluCL. Ibyerekezo bya (A) indole, (B) 4-fluoroindole, (C) 7-iodoindole, na (D) ligands za 5-iodoindole aho GluCL ikora. Poroteyine igaragazwa n'agatambaro, kandi imigozi ya hydrogen y'umugongo igaragazwa nk'imirongo y'umuhondo ifite utudomo. (A′), (B′), (C′), na (D′) bigaragaza imikoranire ya ligands zijyanye n'ibisigazwa bya aside amine biyikikije, kandi imigozi ya hydrogen y'uruhande igaragazwa n'utudomo tw'umutuku.
Igerageza ryakozwe kugira ngo harebwe ingaruka z'uburozi bwa 5-iodoindole ku kumera kw'imbuto z'ibijumba n'amaradishe. 5-iodoindole (0.05 cyangwa 0.1 mM) cyangwa avermectin (10 μg/mL) ntacyo byagize ku kumera kw'imbuto no kumera kw'ibimera (Ishusho ya 7). Byongeye kandi, nta tandukaniro rigaragara ryabonetse hagati y'umuvuduko wo kumera kw'imbuto zitavuwe n'imbuto zavuwe na 5-iodoindole cyangwa avermectin. Ingaruka ku kurekura kw'imizi n'umubare w'imizi yo ku ruhande yari ihari nta gaciro kanini, nubwo 1 mM (inshuro 10 z'umuvuduko wayo) wa 5-iodoindole byatinze gato iterambere ry'imizi yo ku ruhande. Ibi byavuye mu bushakashatsi bigaragaza ko 5-iodoindole idahumanya uturemangingo tw'ibimera kandi ntibangamira iterambere ry'ibimera ku rugero rwakozweho ubushakashatsi.
Ingaruka za 5-iodoindole ku kumera kw'imbuto. Kumera, kumera no gushora imizi ku ruhande rw'imbuto za B. oleracea na R. raphanistrum ku miyoboro ya Murashige na Skoog agar irimo cyangwa idafite avermectin cyangwa 5-iodoindole. Kumera kwanditswe nyuma y'iminsi 3 yo guteka kuri 22°C.
Ubu bushakashatsi bugaragaza ingero nyinshi z’abantu bicwa n’udusimba twitwa indoles. Icy’ingenzi ni uko iyi ari raporo ya mbere yerekana ko iodoindoles itera methylation (inzira iterwa no kwirundanya kwa vacuole nto buhoro buhoro zigahinduka vacuole nini, amaherezo bigatera gucika kw’ururenda no gupfa) mu nshinge za pinusi, aho iodoindoles igaragaza ubushobozi bukomeye bwo kwica udusimba nk’ubwa avermectin yo mu bucuruzi.
Mbere byavuzwe ko Indoles zigira uruhare runini mu gutanga ibimenyetso muri prokaryotes na eukaryotes, harimo gukumira/kwirema kwa biofilm, kubaho kwa bagiteri, no kurwara indwara19,32,33,34. Vuba aha, ingaruka zishobora guterwa n’indoles za halogenated, alkaloids za indoles, n’ibikomoka kuri indoles bya semisynthetic zakuruye ubushakashatsi bwinshi35,36,37. Urugero, indoles za halogenated zagaragaye ko zica uturemangingo twa Escherichia coli na Staphylococcus aureus duhoraho37. Byongeye kandi, ni ingenzi mu bya siyansi kwiga uburyo indoles za halogenated zikora ku zindi moko, amoko, n’ubwami, kandi ubu bushakashatsi ni intambwe igana ku kugera kuri iyi ntego.
Aha, turatanga uburyo bwo kwica 5-iodoindole-induced in C. elegans hashingiwe ku kwangirika kw'uturemangingo (RCI) na methylation (Ishusho ya 4C na 5). Impinduka zo mu mubiri nko kubyimba no kwangirika kwa vacuolar ni ibimenyetso bya RCI na methylation, bigaragazwa nka vacuoles nini muri cytoplasm48,49. RCI ibangamira umusaruro w'ingufu igabanya umusaruro wa ATP, igatera kunanirwa kwa pompe ya ATPase, cyangwa igahungabanya uturemangingo tw'ingirabuzimafatizo bigatuma Na+, Ca2+, na water50,51,52 byinjira vuba. Vacuoles zo mu nda ya cytoplasm zivuka mu turemangingo tw'inyamaswa bitewe n'amazi yirundanyije muri cytoplasm bitewe n'ubwinshi bwa Ca2+ na water53. Igishimishije ni uko ubu buryo bwo kwangirika kw'uturemangingo bushobora gusubizwa inyuma iyo ibyangiritse ari iby'igihe gito kandi uturemangingo tugatangira gukora ATP mu gihe runaka, ariko iyo ibyangiritse bikomeje cyangwa bikarushaho kwiyongera, uturemangingo turapfa.54 Ibyo twabonye bigaragaza ko udusimba twavuwe na 5-iodoindole tudashobora gusubiza imiterere isanzwe y'uturemangingo nyuma yo guhura n'ibibazo by'ingutu.
Uburyo methylation iterwa na 5-iodoindole muri B. xylophilus ishobora guterwa no kuba hari iyode n'uburyo ikwirakwiramo, kubera ko 7-iodoindole yagize ingaruka nke ku B. xylophilus kurusha 5-iodoindole (Imbonerahamwe ya 1 n'Ishusho y'inyongera S6). Ibi bisubizo bihuye gato n'ubushakashatsi bwa Maltese et al. (2014), batangaje ko kwimura moiety ya pyridyl azote muri indole kuva kuri para- kugera kuri meta-position byakuyeho vacuolization, gukumira gukura, no kuvura cytotoxicity mu turemangingo twa U251, bigaragaza ko imikoranire ya molekile n'ahantu runaka muri poroteyine ari ingenzi27,44,45. Imikoranire hagati ya indole cyangwa halogenated indoles na GluCL receptors byagaragaye muri ubu bushakashatsi nabyo bishyigikira iki gitekerezo, kuko 5- na 2-iodoindole byagaragaye ko bifatana na GluCL receptors cyane kurusha izindi indoles zasuzumwe (Ishusho ya 6 n'Ishusho y'inyongera S8). Iyode iri ku mwanya wa kabiri cyangwa uwa gatanu wa indole yagaragaye ko ifatana na leucine 218 ya GluCL receptor binyuze mu migozi ya hydrogen yo mu mugongo, mu gihe izindi indole za halogen na indole ubwabyo bikora imigozi ya hydrogen yo mu ruhande idakomeye hamwe na serine 260 (Ishusho ya 6). Bityo rero, dutekereza ko aho halogen iherereye bigira uruhare runini mu gutuma vacuolar degeneration itera, mu gihe imigozi ya 5-iodoindole ikomeza gufunguka, bityo bigatuma amazi yinjira vuba kandi vacuole igacika. Ariko, uburyo burambuye bwo gukora bwa 5-iodoindole burakomeje kumenyekana.
Mbere yo gukoresha 5-iodoindole mu buryo bufatika, ingaruka z'uburozi bwayo ku bimera zigomba gusesengurwa. Igerageza ryacu ryo kumera kw'imbuto ryagaragaje ko 5-iodoindole nta ngaruka mbi yagize ku kumera kw'imbuto cyangwa inzira zakurikiyeho zo gukura ku gipimo cyakozweho ubushakashatsi (Ishusho ya 7). Bityo, ubu bushakashatsi butanga ishingiro ryo gukoresha 5-iodoindole mu bidukikije kugira ngo hagenzurwe ingaruka mbi z'imiti ya pinusi ku biti bya pinusi.
Raporo zabanjirije iyi zagaragaje ko uburyo bwo kuvura bushingiye kuri indole bushobora kuba uburyo bushoboka bwo gukemura ikibazo cyo kudakira kwa antibiyotike no kwiyongera kwa kanseri55. Byongeye kandi, indole zifite ibikorwa byo kurwanya bagiteri, kurwanya kanseri, antioxidant, kurwanya ububyimbirwe, kurwanya diyabete, kurwanya virusi, kurwanya ikwirakwira ry'imiti no kurwanya igituntu kandi zishobora kuba ishingiro ryiza ryo guteza imbere imiti56,57. Ubu bushakashatsi bwerekana ku nshuro ya mbere ko iyode ishobora gukoreshwa nk'umuti urwanya udukoko n'imiti igabanya ubukana bwa kanseri.
Avermectin yavumbuwe mu myaka mirongo itatu ishize, yegukana igihembo cya Nobel mu 2015, kandi ikoreshwa ryayo nk'umuti ugabanya ubukana riracyakomeje. Ariko, bitewe n'iterambere ryihuse ry'ubudahangarwa bwa avermectin mu mikuyu n'udukoko, hakenewe ubundi buryo, buhendutse kandi butangiza ibidukikije kugira ngo hagenzurwe ubwandu bwa PWN mu biti bya pinusi. Ubu bushakashatsi kandi bugaragaza uburyo 5-iodoindole yica imikuyu n'imikuyu kandi ko 5-iodoindole ifite uburozi buke ku turemangingo tw'ibimera, ibi bikaba bitanga amahirwe meza yo gukoreshwa mu bucuruzi mu gihe kizaza.
Igerageza ryose ryemejwe na Komite Ishinzwe Imyitwarire ya Kaminuza ya Yeungnam, Gyeongsan, muri Koreya, kandi uburyo bwakozwe hakurikijwe amabwiriza ya Komite Ishinzwe Imyitwarire ya Kaminuza ya Yeungnam.
Igerageza ryo gutera amagi mu buryo bwo kuyabyaza umusaruro ryakozwe hakoreshejwe uburyo bwashyizweho43. Kugira ngo harebwe uko avuka (HR), inyange z'umunsi umwe (ingore zigera ku 100 n'ingabo 100) zoherejwe mu biribwa bya Petri birimo fungus maze zirekurwa mu gihe cy'amasaha 24. Hanyuma amagi yakuwe mu yandi maze avurwa na 5-iodoindole (0.05 mM na 0.1 mM) cyangwa avermectin (10 μg/ml) nk'ifatizo mu mazi yasukuwe. Izi fatizo (500 μl; hafi amagi 100) zoherejwe mu mwobo w'isahani y'ibiraro 24 maze zishyirwa kuri 22 °C. Umubare wa L2 wakozwe nyuma y'amasaha 24 yo gutera amagi mu buryo bwo kuyabyaza umusaruro ariko byafatwaga nk'ibyapfuye iyo uturemangingo tutimukaga iyo twakoreshejwe insinga nziza ya platine. Iri gerageza ryakozwe mu byiciro bibiri, buri kimwe kikagira inshuro esheshatu. Amakuru yavuye mu bushakashatsi bwombi yahujwe kandi agaragazwa. Ijanisha rya HR ribazwe ku buryo bukurikira:
Impfu z'ibibyimba zapimwe hakoreshejwe uburyo bwakozwe mbere. Amagi ya Nematode yakusanyijwe hanyuma insoro zihuzwa no gukururwa mu mazi yanduye kugira ngo habeho ibibyimba bya L2. Ibibyimba byahuje (hafi ibibyimba 500) byavuwe na 5-iodoindole (0.05 mM na 0.1 mM) cyangwa avermectin (10 μg/ml) maze birererwa ku mabati ya B. cinerea Petri. Nyuma y'amasaha 48 yo gutwikirwa kuri 22 °C, ibibyimba byakusanyijwe mu mazi yanduye maze bisuzumwa niba hari ibyiciro bya L2, L3, na L4. Kuba hari ibyiciro bya L3 na L4 byagaragaje ko ibibyimba byahindutse, mu gihe kuba hari icyiciro cya L2 byagaragaje ko nta mpinduka zahindutse. Amashusho yabonetse hakoreshejwe Sisitemu ya iRiS™ Digital Cell Imaging System. Iri gerageza ryakozwe mu byiciro bibiri, buri kimwe kikagira inshuro esheshatu. Amakuru yavuye mu bushakashatsi bwombi yahujwe kandi yerekanwa.
Uburozi bwa 5-iodoindole na avermectin ku mbuto bwapimwe hakoreshejwe ibizamini byo kumera ku masahani ya Murashige na Skoog agar.62 Imbuto za B. oleracea na R. raphanistrum zabanje kwinjizwa mu mazi yanduye mu gihe cy'umunsi umwe, zozwa na ml 1 ya ethanol 100%, zishyirwamo bleach ya 50% yo mu bucuruzi (3% sodium hypochlorite) mu gihe cy'iminota 15, hanyuma zozwa inshuro eshanu na ml 1 y'amazi yanduye. Imbuto zanduye zashyizwe ku masahani ya agar yo mu burumbuke arimo 0.86 g/l (0.2X) Murashige na Skoog medium na 0.7% bacteriological agar irimo cyangwa idafite 5-iodoindole cyangwa avermectin. Hanyuma amasahani yashyizwe ku bushyuhe bwa 22 °C, maze amashusho afatwa nyuma y'iminsi 3 yo kwinjizwa. Iri gerageza ryakozwe mu byiciro bibiri, buri kimwe muri byo kikaba cyarasubiwemo inshuro esheshatu.
Igihe cyo kohereza: Gashyantare-26-2025