Ngā rauemi tipu me ngā matūkita
I whakaratohia e Tākuta Pat Brown i Te Whare Wānanga o Illinois (kei UC Davis ināianei) tētahi taupori mahere hononga sorghum e mōhiotia ana ko te taupori huringa sorghum (SCP). Kua whakaahuahia i mua ake nei, ā, he kohinga o ngā momo momo kua hurihia kia kore e aro ki te wā-whakaahua me te iti ake te roa hei whakahaere i te tipu me te whanaketanga o ngā tipu i ngā taiao o Amerika. E 510 ngā momo momo mai i tēnei taupori i whakamahia i roto i tēnei rangahau, ahakoa nā te kino o te whakatipu me ētahi atu take whakahaere kounga, kāore i whakamahia ngā momo katoa i roto i te tātari i ngā āhuatanga e toru. I te mutunga, i whakamahia ngā raraunga mai i ngā momo 345 hei tātari i te urupare chitin, 472 ngā momo mō te urupare flg22, me te 456 mō te ātete TLS.B. kukiiI riro mai te momo LSLP18 i a Tākuta Burt Bluhm i Te Whare Wānanga o Arkansas.
Te ine i te urupare MAMP
E rua ngā MAMP rerekē i whakamahia i roto i tēnei rangahau flg22, (Genscript catalog# RP19986), me te chitin. I whakatipuria ngā tipu Sorghum i roto i ngā whakaurunga i whakatakotoria ki runga i ngā papatahi kua whakakiia ki te oneone (33% Sunshine Redi-Earth Pro Growing Mix) i roto i te whare kati kōtuhi. I whakamakukuhia ngā tipu i te rā i mua i te kohikohinga tauira hei karo i te makuku rau i te rā o te kohikohinga.
I whakarōpūtia matapōkeretia ngā rārangi, ā, mō ngā take whakarite, i whakatōkia ki roto i ngā puranga o te 60 rārangi. Mō ia rārangi, e toru ngā 'ipu' i whakatōkia me ngā kākano e rua mō ia rārangi. I whakatōkia ngā puranga i muri mai i te tukatuka o te puranga o mua tae noa ki te aromatawai i te taupori katoa. E rua ngā whakamātautau i whakahaerehia mō ngā MAMP e rua me ngā iraira i whakarōpūtia matapōkeretia anō i ia o ngā oma e rua.
I whakahaerehia ngā whakamātautau ROS e ai ki te whakaahuatanga o mua. I roto i te poto, mō ia rārangi, e ono ngā kākano i whakatōkia ki roto i ngā kohua rerekē e toru. Mai i ngā tipu i puta mai, e toru i tīpakohia i runga i te ōritetanga. Kāore i whakamahia ngā tipu i āhua rerekē, i teitei ake, i poto ake rānei i te nuinga. E whā ngā kōpae rau, 3 mm te whānui, i tangohia mai i te wāhanga whānui rawa o te rau tuawhā o ngā tipu sorghum rerekē e toru, 15-rā te pakeke. Kotahi te kōpae mō ia rau mai i ngā tipu e rua, e rua hoki ngā kōpae mai i te tipu kotahi, ko te kōpae tuarua hei mana whakahaere wai (tirohia i raro). I whakamānuhia takitahitia ngā kōpae ki runga i te 50 µl H20 i roto i tētahi pereti pango 96-puna, i hiritia ki te hiri konumohe hei karo i te pānga ki te mārama, ā, i puritia ki te pāmahana rūma mō te pō. I te ata i muri mai, i hangaia he otinga tauhohenga mā te whakamahi i te 2 mg/ml probe chemiluminescent L-012 (Wako, putumōhio # 120-04891), 2 mg/ml horseradish peroxidase (Momo VI-A, Sigma-Aldrich, putumōhio # P6782), me te 100 mg/ml Chitin, 2 μM rānei o Flg22. I tāpirihia he 50 µl o tēnei otinga tauhohenga ki ngā puna e toru o ngā puna e whā. Ko te puna tuawhā he mana whakahaere tauira, ā, i tāpirihia te otinga tauhohenga kāore i te whakaurua te MAMP. E whā ngā puna kau kei roto ko te wai anake i whakaurua ki ia pereti.
I muri i te tāpiri i te otinga tauhohenga, i inehia te kanapa mā te whakamahi i te kaipānui pereti maha-kitenga SynergyTM 2 (BioTek) i ia 2 meneti mō te 1 hāora. Ka inehia te kanapa e te kaipānui pereti i ia 2 meneti i roto i tēnei 1 hāora. I tatauhia te tapeke o ngā pānuitanga 31 katoa hei homai i te uara mō ia puna. I tatauhia te uara whakatau tata mō te urupare MAMP mō ia momoira hei (te uara kanapa toharite o ngā puna whakamātautau e toru—te uara puna tauira) -engari kāore te uara puna kau noa. I tata tonu ngā uara puna kau noa ki te kore.
Ngā kōpae rau oNicotiana benthamiana, kotahi te raina sorghum aro nui (SC0003), me tetahi raina sorghum aro iti (PI 6069) i whakaurua hoki hei mana whakahaere i roto i ia pereti 96-poka mō ngā kaupapa whakahaere kounga.
B. kukiite whakarite i te werohanga me te werohanga
B. kukiiI whakaritea te werohanga pēnei i te whakaahuatanga o mua. I roto i te poto, i rumakina ngā pata sorghum ki te wai mō ngā rā e toru, i horoia, i kokohia ki roto i ngā ipu kōniko 1L, ā, i whakamahanatia mō te haora i te 15psi me te 121 °C. Kātahi ka werohia ngā pata ki te 5 ml o te mycelia kua whakamaroketia mai i tētahi ahurea hou oB. kukiiKa tangohia ngā momo LSLP18, ka waiho mō ngā wiki e rua i te pāmahana rūma, me te rūrū i ngā ipu i ia rā e toru. I muri i ngā wiki e rua, ka whakamaroketia ngā pata sorghum kua pangia e te harore ki te hau, kātahi ka rongoatia ki te 4 °C tae noa ki te werohanga i te mara. I whakamahia te werohanga kotahi mō te whakamātautau katoa, ā, i hangaia kia hou i ia tau. Mō te werohanga, ka whakanohoia kia 6–10 ngā pata kua pangia ki roto i te porowhita o ngā tipu sorghum 4–5 wiki te pakeke. Nā ngā pua i puta mai i ēnei harore i tīmata te mate ki ngā tipu sorghum taitamariki i roto i te wiki kotahi.
Te whakarite purapura
I mua i te whakatō ki te mara, i rongoāhia te purapura sorghum ki te harore patu, te patupatu pepeke, me te ranunga safener kei roto ko te ~ 1% patupatu Spirato 480 FS, 4% patupatu Sebring 480 FS, 3% Sorpro 940 ES purapura safener. Kātahi ka whakamaroketia ngā purapura ki te hau mō ngā rā e 3, kia kapi ai ngā purapura i te ranunga. Nā te safener i taea ai te whakamahi i te patupatu pepeke Dual Magnum hei rongoā i mua i te puta mai.
Aromatawai i te ātete ki ngā ira rau whāinga
I whakatōkia te SCP ki te Teihana Rangahau Hua Matua i Clayton, NC i te 14–15 o Hune 2017 me te 20 o Hune, 2018 i roto i tētahi hoahoa poraka matapōkere katoa me ngā tukuruatanga whakamātautau e rua i ia kēhi. I whakatōkia ngā whakamātautau ki ngā rarangi takitahi 1.8 m me te whānui rarangi 0.9 m mā te whakamahi i te 10 purapura mō ia poraka. E rua ngā rarangi rohe i whakatōkia ki te taha o ia whakamātautau hei ārai i ngā pānga taha. I werohia ngā whakamātautau i te 20 o Hūrae, 2017 me te 20 o Hūrae, 2018, i taua wā kua tae ngā tipu sorghum ki te wāhanga tipu 3. I tangohia ngā whakatauranga i runga i ngā tauine kotahi ki te iwa, i reira ko ngā tipu kāore e whakaatu ana i ngā tohu o te mate i tohua hei iwa, ā, ko ngā tipu kua mate rawa i tohua hei kotahi. E rua ngā whakatauranga i tangohia i te tau 2017, ā, e whā ngā pānui i te tau 2018 e tīmata ana i ngā wiki e rua i muri i te werohanga i ia tau. I tatauhia te sAUDPC (whare paerewa i raro i te pihi haere whakamua o te mate) e ai ki te whakaahuatanga o mua.
Wā tuku: Paenga-whāwhā-01-2021