Nā mea kanu a me nā mea hoʻomake
Ua hāʻawi ʻia kahi heluna kanaka hoʻololi sorghum (SCP) e Kauka Pat Brown ma ke Kulanui o Illinois (i kēia manawa ma UC Davis). Ua wehewehe mua ʻia a he hōʻiliʻili o nā laina like ʻole i hoʻololi ʻia i ka photoperiod-insensitivity a me ke kūlana liʻiliʻi e hoʻomaʻamaʻa i ka ulu ʻana a me ka hoʻomohala ʻana o nā mea kanu ma nā wahi US. Ua hoʻohana ʻia he 510 mau laina mai kēia heluna kanaka ma kēia haʻawina ʻoiai ma muli o ka germination maikaʻi ʻole a me nā pilikia kaohi maikaʻi ʻē aʻe, ʻaʻole i hoʻohana ʻia nā laina āpau i ka nānā ʻana i nā ʻano ʻekolu. I ka hopena, ua hoʻohana ʻia nā ʻikepili mai 345 mau laina no ka nānā ʻana i ka pane chitin, 472 mau laina no ka pane flg22, a me 456 no ke kūʻē ʻana o TLS.B. kukiUa loaʻa ke ʻano LSLP18 mai Kauka Burt Bluhm ma ke Kulanui o Arkansas.
Ana pane MAMP
Ua hoʻohana ʻia ʻelua mau MAMP like ʻole ma kēia noiʻi flg22, (Genscript catalog# RP19986), a me chitin. Ua ulu ʻia nā mea kanu Sorghum i loko o nā mea i hoʻokomo ʻia ma nā pālahalaha i piha i ka lepo (33% Sunshine Redi-Earth Pro Growing Mix) i loko o ka hale kōmahana. Ua hoʻoinu ʻia nā mea kanu i ka lā ma mua o ka hōʻiliʻili ʻana i ka laʻana e pale aku i ka makū lau keu i ka lā o ka hōʻiliʻili ʻana.
Ua koho wale ʻia nā laina a, no nā kumu logistical, ua kanu ʻia i loko o nā pūʻulu o 60 mau laina. No kēlā me kēia laina, ua kanu ʻia ʻekolu 'ipu' me ʻelua mau hua no kēlā me kēia laina. Ua kanu ʻia nā pūʻulu ma hope koke iho o ka hana ʻia ʻana o ka pūʻulu ma mua a hiki i ka loiloi ʻia ʻana o ka heluna kanaka holoʻokoʻa. Ua mālama ʻia ʻelua mau holo hoʻokolohua no nā MAMP ʻelua me nā genotypes i koho hou ʻia i kēlā me kēia o nā holo ʻelua.
Ua hoʻokō ʻia nā hoʻāʻo ROS e like me ka mea i wehewehe mua ʻia. I ka pōkole, no kēlā me kēia laina, ua kanu ʻia ʻeono mau hua i loko o 3 mau ipu like ʻole. Mai nā mea kanu i loaʻa mai, ua koho ʻia ʻekolu ma muli o ke ʻano like. ʻAʻole i hoʻohana ʻia nā mea kanu i ʻano ʻē a i ʻole he kiʻekiʻe loa a pōkole paha ma mua o ka hapa nui. Ua ʻoki ʻia ʻehā mau diski lau o 3 mm ke anawaena mai ka ʻāpana ākea loa o ka lau ʻehā o ʻekolu mau mea kanu sorghum 15 lā like ʻole. Hoʻokahi diski no kēlā me kēia lau mai ʻelua mau mea kanu a ʻelua mau diski mai hoʻokahi mea kanu, me ka lua o ka diski e lilo i ka mana wai (e nānā ma lalo). Ua lana pākahi ʻia nā diski ma luna o 50 µl H20 i loko o kahi pā ʻeleʻele 96-lua, i hoʻopaʻa ʻia me kahi sila alumini e pale aku i ka ʻike ʻia ʻana i ka mālamalama, a mālama ʻia i ka mahana o ka lumi i ka pō. I ke kakahiaka aʻe, ua hana ʻia kahi hopena hopena me ka hoʻohana ʻana i ka 2 mg/ml chemiluminescent probe L-012 (Wako, catalog # 120-04891), 2 mg/ml horseradish peroxidase (ʻAno VI-A, Sigma-Aldrich, catalog # P6782), a me 100 mg/ml Chitin a i ʻole 2 μM o Flg22. Ua hoʻohui ʻia he 50 µl o kēia hopena hopena i ʻekolu o nā lua ʻehā. ʻO ka lua ʻehā he mana hoʻohālike, kahi i hoʻohui ʻia ai ka hopena hopena me ka ʻole o ka MAMP. Ua hoʻokomo pū ʻia ʻehā mau lua hakahaka me ka wai wale nō i loko o kēlā me kēia pā.
Ma hope o ka hoʻohui ʻana i ka hopena hopena, ua ana ʻia ka luminescence me ka hoʻohana ʻana i ka SynergyTM 2 multi-detection microplate reader (BioTek) i kēlā me kēia 2 min no 1 hola. Lawe ka mea heluhelu papa i nā ana luminescence i kēlā me kēia 2 min i loko o kēia 1 hola. Ua helu ʻia ka huina o nā heluhelu 31 āpau e hāʻawi i ka waiwai no kēlā me kēia lua. Ua helu ʻia ka waiwai i manaʻo ʻia no ka pane MAMP no kēlā me kēia genotype e like me (ka awelika o ka waiwai luminescence o nā lua hoʻokolohua ʻekolu—ka waiwai lua hoʻohenehene) -me ka awelika o ka waiwai lua hakahaka. Ua kokoke mau nā waiwai lua hakahaka i ka ʻole.
Nā diski lau oʻO Nicotiana benthamiana, hoʻokahi laina sorghum pane kiʻekiʻe (SC0003), a hoʻokahi laina sorghum pane haʻahaʻa (PI 6069) i hoʻokomo pū ʻia ma ke ʻano he mau kaohi i kēlā me kēia pā 96-lua no nā kumu kaohi maikaʻi.
B. kukika hoʻomākaukau ʻana i ka inoculum a me ka inoculation
B. kukiUa hoʻomākaukau ʻia ka inoculum e like me ka mea i wehewehe mua ʻia. I ka pōkole, ua hoʻomoʻa ʻia nā hua sorghum i ka wai no ʻekolu lā, holoi ʻia, ʻeli ʻia i loko o nā ʻōmole conical 1L a hoʻomoʻa ʻia i loko o ka autoclave no hoʻokahi hola ma 15psi a me 121 °C. A laila ua hoʻokomo ʻia nā hua me kahi 5 ml o ka mycelia macerated mai kahi moʻomeheu hou oB. kukiHoʻokaʻawale ʻo LSLP18 a waiho ʻia no 2 mau pule ma ka mahana o ka lumi, e lūlū ana i nā ʻōmole i kēlā me kēia 3 lā. Ma hope o 2 mau pule, ua hoʻomaloʻo ʻia nā hua sorghum i piha i ka fungus i ka ea a laila mālama ʻia ma 4 °C a hiki i ka hoʻopili ʻana i ke kula. Ua hoʻohana ʻia ka inoculum like no ka hoʻokolohua holoʻokoʻa a hana hou ʻia i kēlā me kēia makahiki. No ka hoʻopili ʻana, ua kau ʻia he 6-10 mau hua i piha i ka whorl o nā mea kanu sorghum 4-5 mau pule. ʻO nā spores i hana ʻia mai kēia mau fungi i hoʻomaka i ka maʻi i loko o nā mea kanu sorghum ʻōpio i loko o hoʻokahi pule.
Hoʻomākaukau hua
Ma mua o ke kanu ʻana ma ke kula, ua mālama ʻia ka hua sorghum me kahi fungicide, insecticide, a me ka hui safener e loaʻa ana he ~ 1% Spirato 480 FS fungicide, 4% Sebring 480 FS fungicide, 3% Sorpro 940 ES seed safener. A laila ua hoʻomaloʻo ʻia nā hua i ka ea no 3 mau lā i hāʻawi i kahi uhi lahilahi o kēia hui a puni nā hua. Ua ʻae ka safener i ka hoʻohana ʻana i ka herbicide Dual Magnum ma ke ʻano he lāʻau lapaʻau ma mua o ka puka ʻana mai.
Ka loiloi ʻana o ke kū'ē ʻana o ka Target Leaf Spot
Ua kanu ʻia ka SCP ma ke Keʻena Noiʻi Meakanu Waena ma Clayton, NC ma Iune 14-15 2017 a me Iune 20, 2018 ma kahi hoʻolālā poloka piha randomized me ʻelua mau hana hou hoʻokolohua ma kēlā me kēia hihia. Ua kanu ʻia nā hoʻokolohua ma nā lālani hoʻokahi he 1.8 m me ka laulā lālani 0.9 m me ka hoʻohana ʻana i 10 mau hua no kēlā me kēia ʻāpana. Ua kanu ʻia ʻelua mau lālani palena a puni ka periphery o kēlā me kēia hoʻokolohua e pale aku i nā hopena lihi. Ua hoʻokomo ʻia nā hoʻokolohua ma Iulai 20, 2017 a me Iulai 20, 2018 ma kahi manawa i loaʻa ai nā mea kanu sorghum i ke kahua ulu 3. Ua lawe ʻia nā helu ma kahi unahi hoʻokahi a ʻeiwa, kahi i helu ʻia ai nā mea kanu e hōʻike ʻole ana i nā hōʻailona o ka maʻi ma ke ʻano he ʻeiwa a ua helu ʻia nā mea kanu make loa ma ke ʻano he hoʻokahi. Ua lawe ʻia ʻelua mau helu i ka makahiki 2017 a me ʻehā mau heluhelu i ka makahiki 2018 e hoʻomaka ana i ʻelua pule ma hope o ka hoʻokomo ʻana i kēlā me kēia makahiki. Ua helu ʻia ʻo sAUDPC (standardized area under disease progression curve) e like me ka mea i wehewehe mua ʻia.
Ka manawa hoʻouna: ʻApelila-01-2021