Ana iya amfani da naman gwari mai fashewa na shinkafa Kosakonia oryziphila NP19 a matsayin mai haɓaka haɓakar shuka da kuma maganin kashe kwari na halitta don rage fashewar shinkafa na nau'in KDML105.

Wannan binciken ya nuna cewa naman gwari mai suna rhizosphere symbiotic *Kosakonia oryziphila* NP19 da aka ware daga tushen shinkafa wani kyakkyawan maganin kashe kwari ne da ke haɓaka haɓakar shuka don magance fashewar shinkafa da *Pyricularia oryzae* ke haifarwa. An gudanar da gwaje-gwaje a cikin vitro akan sabbin ganyen shinkafa jasmine na nau'in Khao Dawk Mali 105 (KDML105). Sakamakon ya nuna cewa NP19 ya hana haɓakar *Pyricularia oryzae* conidia yadda ya kamata. An hana kamuwa da cutar *Pyricularia oryzae* a ƙarƙashin yanayi uku daban-daban na magani: na farko, an yi wa shinkafar da NP19 kuma an yi mata allurar *Pyricularia oryzae* conidia; na biyu, an shafa cakuda NP19 da *Pyricularia oryzae* conidia a cikin ganyayyaki;
Kwayoyin cutar rhizosphere *Kosakonia oryziphila* NP19¹⁴an ware shi daga tushen shinkafa (*Oryza sativa* L. cv. RD6). *Kosakonia oryziphila* NP19 yana da kaddarorin haɓaka haɓakar shuka, gami da daidaita nitrogen, samar da indoleacetic acid (IAA), da kuma narkewar phosphate. Abin sha'awa, *Kosakonia oryziphila* NP19 yana samar da chitinase^14.Amfani da *Kosakonia oryziphila* NP19 zuwa KDML105 iri shinkafa ya inganta rayuwar shinkafa bayan kamuwa da fashewar shinkafa. Manufar wannan binciken ita ce (i) bayyana hanyar hana *Kosakonia oryziphila* NP19 akan fashewar shinkafa da kuma (ii) bincika tasirin *Kosakonia oryziphila* NP19 wajen sarrafa fashewar shinkafa.

Sinadaran gina jiki suna taka muhimmiyar rawa wajen girma da ci gaban shuka, suna aiki a matsayin abubuwan da ke sarrafa cututtuka daban-daban na ƙwayoyin cuta. Abincin ma'adinai na shuka yana ƙayyade juriyarsa ga cututtuka, halayen siffa ko nama, da kuma ƙarfinsa, ko ikon rayuwa daga ƙwayoyin cuta. Phosphorus na iya rage ci gaban da rage tsananin fashewar shinkafa ta hanyar ƙara haɗakar mahaɗan phenolic. Potassium gabaɗaya yana rage yawan kamuwa da cututtukan shinkafa da yawa, kamar fashewar shinkafa, tabo na ganyen bakteriya, tabo na ganye, ruɓewar tushe, da tabo na ganye. Wani bincike da Perrenoud ya yi ya nuna cewa takin mai yawan potassium na iya rage yawan kamuwa da cututtukan fungal na shinkafa da ƙara yawan amfanin gona. Nazari da yawa sun nuna cewa takin sulfur na iya inganta juriyar amfanin gona ga ƙwayoyin fungal.²⁷Yawan sinadarin magnesium (wani sinadari na chlorophyll) na iya haifar da fashewar shinkafa.²¹Zinc zai iya kashe ƙwayoyin cuta kai tsaye, ta haka yana rage tsananin cutar.²²Gwaje-gwajen da aka gudanar a gona sun nuna cewa duk da cewa yawan phosphorus, potassium, sulfur, da zinc a cikin ƙasa ya fi na gwajin tukunya yawa, fashewar shinkafa har yanzu tana yaɗuwa ta cikin ganyen shinkafa. Sinadaran ƙasa ba za su yi tasiri sosai wajen sarrafa fashewar shinkafa ba, domin ɗanɗano da zafin jiki ba su da kyau ga kamuwa da ƙwayoyin cuta masu ƙarfi.
A gwaje-gwajen da aka yi a filin, an gano Stenotrophomonas maltophilia, P. dispersa, Xanthomonas sacchari, Burkholderia multivorans, Burkholderia diffusa, Burkholderia vietnamiensis da C. gleum a duk hanyoyin magani. An ware Stenotrophomonas maltophilia daga rhizosphere na alkama, hatsi, kokwamba, masara, da dankali kuma ya nuna ikon sarrafa halittu.^aikia kan Colletotrichum nymphaeae.28 Bugu da ƙari, an ruwaito cewa P. dispersa yana da tasiri a kan baƙi^ruɓarDankali mai zaki.29 Bugu da ƙari, nau'in R1 na Xanthomonas sacchari ya nuna ƙarfin hali game da fashewar shinkafa da ruɓewar panicle da Burkholderia ke haifarwa.^glumae.30Burkholderia oryzae NP19 na iya ƙulla alaƙar symbiosis da ƙwayar shinkafa yayin tsiro kuma ya zama naman gwari mai kama da na wasu nau'ikan shinkafa. Yayin da wasu ƙwayoyin cuta na ƙasa za su iya mamaye shinkafa bayan dasawa, naman gwari mai kama da na NP19, da zarar ya zama mamaya, yana shafar abubuwa da yawa a cikin tsarin kariya na shinkafa daga wannan cuta. NP19 ba wai kawai yana hana ci gaban P. oryzae da fiye da 50% ba (duba Tebur na Ƙarin S1 a cikin ƙarin bayani na kan layi), amma kuma yana rage adadin raunukan fashewa a kan ganye kuma yana ƙara yawan amfanin shinkafar da aka yi wa allurar rigakafi ko aka mamaye ta da NP19 (RBf, RFf-B, da RBFf-B) a cikin gwaje-gwajen gona (Hoto na S3).
Naman gwari na Pyricularia oryzae, wanda ke haifar da fashewar tsirrai, naman gwari ne mai hemittrophic wanda ke buƙatar abubuwan gina jiki daga shukar da ke karɓar baƙi yayin kamuwa da cuta. Tsire-tsire suna samar da nau'in iskar oxygen mai amsawa (ROS) don rage kamuwa da cutar fungal; duk da haka, Pyricularia oryzae yana amfani da dabaru daban-daban don magance ROS da mai karɓar baƙi ke samarwa.³¹Da alama Peroxidases suna taka rawa wajen juriya ga ƙwayoyin cuta, ciki har da haɗakar sunadaran bangon tantanin halitta, kauri ganuwar xylem, samar da ROS, da kuma kawar da hydrogen peroxide.³²Sinadaran hana tsufa na iya zama wani takamaiman tsarin gano ROS. Ta hanyar halayensu na hana tsufa, superoxide dismutase (SOD) da peroxidase (POD) suna taimakawa wajen fara martanin kariya, tare da SOD suna aiki a matsayin layin farko na kariya.³³A cikin shinkafa, ana haifar da aikin peroxidase na shuka bayan kamuwa da ƙwayoyin cuta na shuka kamar *Pyricularia oryzae* da *Xanthomonas oryzae pv. Oryzae*.³²A cikin wannan binciken, aikin peroxidase ya ƙaru a cikin shinkafar da aka mamaye ko kuma/ko aka yi wa allurar *Magnaporthe oryzae* NP19; duk da haka, *Magnaporthe oryzae* bai shafi aikin peroxidase ba. Superoxide dismutase (SOD), kamar yadda H₂O₂ synthase, ke haifar da raguwar O₂⁻ zuwa H₂O₂. SOD yana taka muhimmiyar rawa wajen juriyar tsirrai ga matsaloli daban-daban ta hanyar daidaita yawan H₂O₂ a cikin shukar, ta haka yana haɓaka juriyar shuka ga matsaloli daban-daban³⁴. A cikin wannan binciken, a cikin gwajin tukunya, kwanaki 30 bayan allurar *Magnaporthe oryzae* (30 DAT), ayyukan SOD a cikin ƙungiyoyin RF da RBF sun kasance 121.9% da 104.5% fiye da waɗanda ke cikin rukunin R, bi da bi, suna nuna martanin SOD ga kamuwa da cutar *Magnaporthe oryzae*. A gwaje-gwajen tukunya da na fili, ayyukan SOD a cikin shinkafar da aka yi wa allurar *Magnaporthe oryzae* NP19 sun fi na shinkafar da ba a yi wa allurar ba da kashi 67.7% da 28.8% fiye da ta shinkafar da ba a yi wa allurar ba kwanaki 30 bayan allurar, bi da bi. Amsoshin sinadarai na tsirrai suna shafar muhalli, tushen damuwa, da nau'in shuka³⁵. Ayyukan enzyme na antioxidant na shuka suna shafar kai tsaye ta hanyar abubuwan muhalli, waɗanda kuma ke shafar ayyukan enzyme na antioxidant na shuka ta hanyar canza al'ummar ƙwayoyin cuta na shuka.
An yi amfani da wannan binciken a matsayin nau'in fungi na cutar busasshen shinkafa (Kosakonia oryziphila NP19, lambar shiga ta NCBI PP861312)¹³An ware shi daga tushen nau'in shinkafa RD6 a Lardin Nakhon Phanom, Thailand (16° 59′ 42.9″ N 104° 22′ 17.9″ E). An noma wannan nau'in a cikin ruwan sinadirai (NB) a zafin 30°C da 150 rpm na tsawon awanni 18. Don ƙididdige yawan ƙwayoyin cuta, an auna shan dakatarwar ƙwayoyin cuta a 600 nm. An daidaita yawan dakatarwar ƙwayoyin cuta zuwa^10⁶CFU/mL tare da ruwan da aka cire ion (_dH₂O). An yi wa naman gwari mai fashewa (Pyricularia oryzae) allurar tabo a kan dankalin turawa dextrose agar (PDA) sannan aka saka shi a zafin 25°C na tsawon kwanaki 7. An mayar da mycelium na naman gwari zuwa matsakaicin bran bran (2% (w/v) bran shinkafa, 0.5% (w/v) sucrose, da 2% (w/v) agar da aka narkar a cikin ruwan da aka cire ion, pH 7) sannan aka saka shi a zafin 25°C na tsawon kwanaki 7. An sanya ganyen da aka tsaftace na wani nau'in shinkafa mai saurin kamuwa da cuta (KDML105) a kan mycelium don haifar da conidia kuma aka saka shi a zafin 25°C na tsawon kwanaki 5 a ƙarƙashin hasken UV da fari. An tattara Conidia ta hanyar goge mycelium da saman ganyen da ya kamu da cutar a hankali da 10 ml na maganin 0.025% (v/v) Tween 20. An tace maganin fungal ta hanyar yadudduka takwas na cheesecloth don cire mycelium, agar, da ganyen shinkafa. An daidaita yawan sinadarin conidia a cikin dakatarwar zuwa 5 × 10⁵ conidia/ml don ƙarin bincike.
An shirya sabbin al'adun ƙwayoyin Kosakonia oryziphila NP19 ta hanyar noma su a cikin matsakaicin NB a zafin jiki na 37 °C na tsawon awanni 24. Bayan an yi amfani da centrifugation (3047 × g, minti 10), an tattara ƙwayoyin ƙwayoyin, an wanke su sau biyu da 10 mM phosphate-buffered saline (PBS, pH 7.2), sannan aka sake dakatar da su a cikin wannan buffer. An auna yawan gani na dakatarwar ƙwayoyin a 600 nm, inda aka sami ƙimar kusan 1.0 (daidai da 1.0 × 10⁷ CFU/μl wanda aka ƙayyade ta hanyar shafa su a kan faranti na agar na gina jiki). An sami Conidia na P. oryzae ta hanyar dakatar da su a cikin maganin PBS da ƙidaya su ta amfani da hemocytometer. Dakatarwar *K. oryziphila* NP19 da *P. Don gwaje-gwajen shafawa na ganye, an shirya K. oryziphila* conidia a kan sabbin ganyen shinkafa a yawan 1.0 × 10⁷ CFU/μL da 5.0 × 10² conidia/μL, bi da bi. Hanyar shirya samfurin shinkafar ita ce kamar haka: An yanke ganyen shinkafa mai tsawon santimita 5 kuma an sanya su a cikin kwano na Petri da aka lulluɓe da takarda mai ɗanɗano. An kafa ƙungiyoyi biyar na magani: (i) R: ganyen shinkafa ba tare da allurar ƙwayoyin cuta ba a matsayin magani, an ƙara musu 0.025% (v/v) maganin Twin 20; (ii) RB + F: an yi wa shinkafa allurar K. oryziphila NP19, an ƙara musu 2 μL na dakatarwar conidia na naman gwari wanda ke haifar da fashewar shinkafa; (iii) R + BF: Shinkafa a cikin rukunin R an ƙara musu 4 μl na cakuda dakatarwar fungal conidia da K. oryziphila NP19 (rabo mai girma 1:1); (iv) R + F: Shinkafa a cikin rukunin R an ƙara ta da 2 μl na blast fungal conidia dakatarwa; (v) RF + B: Shinkafa a cikin rukunin R an ƙara ta da 2 μl na blast fungal conidia dakatarwa na tsawon awanni 30, sannan aka ƙara 2 μl na K. oryziphila NP19 a wuri ɗaya. An saka duk abincin Petri a zafin jiki na 25°C a cikin duhu na tsawon awanni 30 sannan aka sanya su a ƙarƙashin haske mai ci gaba. An samar da kowace rukuni a cikin sau uku. Bayan awanni 72 na noma, an lura da kyallen shukar kuma an yi nazari a kansu ta hanyar duba na'urar daukar hoto ta electron microscopy (SEM). A takaice, an sanya kyallen shukar a cikin phosphate buffer wanda ke ɗauke da 2.5% (v/v) glutaraldehyde kuma an fitar da su ta hanyar jerin maganin ethanol. Bayan bushewar ma'auni mai mahimmanci da carbon dioxide, an shafa samfuran da zinare kuma a ƙarshe an duba su ta amfani da na'urar daukar hoto ta electron scanning.¹⁵