KOLOKOLO MICROBIAL
ʻO ka hoʻoikaika ʻana i ka mahiʻai e hōʻemi i ka paʻakikī o ka ʻoihana microbial a me ka nui o ka ʻauhau kīwī ma nā aʻa
ʻO ke kaʻina Amplicon Sequencing (ITS) |PacBio HIFI
Nā mea nui
AUa lilo ka hoʻoikaika ʻana i ka mahiʻai i ka pilikia ma muli o kona mau hopena ʻino i ke kaiapuni e like me ka maikaʻi ʻole o ka hoʻohana ʻana i nā meaʻai, ka eutrophiation wai gound, degradation o ka maikaʻi o ka lepo, a me nā mea ʻē aʻe.He kuleana koʻikoʻi ke kaiāulu microbial i ka huahana a me ka hoʻomau o ka agroecosystem.Akā naʻe, ʻaʻole maopopo i ka hopena o nā ʻōnaehana mahiʻai ʻokoʻa i ka microbiota aʻa.
Hoʻolālā hoʻokolohua
Nā hoʻokolohua
SʻO nā ʻaila a me ke kumu (DNA) mai nā māla palaoa mai 60 mau ʻāina mahiʻai (20 kēlā me kēia)
Grouping: 1. Convention (with tillage);2. Convention (no-tilage);3. ʻĀina mahiʻai
Shoʻolālā equencing: Full-longth amplicon sequencing (ITS)
Primers: ITS1F-ITS4 (e huli ana i ka māhele ITS holoʻokoʻa ~630 bp)
Skahua hoʻohālikelike: PacBio RS II
ʻIkepili Bioinformatic
Nā hualoaʻa
On ka awelika o 357 OTU i ike ia ma ka paena a me ka huina o 837 OTU o na wahi 60 a pau.ʻAʻole ʻike ʻia ka ʻokoʻa koʻikoʻi o ka ʻokoʻa Alpha o nā kaiāulu aʻa ma waena o nā ʻōnaehana mahiʻai ʻekolu.Eia naʻe, ua hoʻokumu ʻia ʻekolu pūʻulu ʻokoʻa i ka nānā ʻana i nā ʻano like ʻole beta, e hōʻike ana i ka hopena ikaika o ka ʻōnaehana mahiʻai ma ke ʻano o ke kaiaulu fungal root.
Kiʻi 1. ʻano like ʻole Alpha (Shannon index and community composition) a me beta diversity analysis (canonical analysis of principal coordinates) ma nā kaiaulu fungal root.
TUa wehewehe ʻia ka ʻauhau ʻauhau ma muli o ka pūnaewele holoʻokoʻa o nā kaiāulu fungal ma waena o ʻekolu ʻōnaehana mahiʻai: koho ʻia nā nodes 10 kiʻekiʻe me ke kiʻekiʻe kiʻekiʻe, ke kikowaena pili kiʻekiʻe a me ke kikowaena waena haʻahaʻa i koho ʻia.ʻEhiku o lākou no nā kauoha mycorrhizal.
Kiʻi 2. Pūnaewele holoʻokoʻa ma nā kaiaulu fungal aʻa o nā ʻōnaehana mahiʻai ʻekolu
FUa hōʻike ʻia nā ʻupena kikoʻī ʻo arming-system i kahi pilina ʻoi aku ka kiʻekiʻe o ka pūnaewele organik me ʻelua mau ʻaoʻao a ʻoi aku ka nui o nā node i hoʻopili ʻia ma mua o ka pūnaewele no-till a maʻamau.Eia kekahi, ʻoi aku ka nui o ka ʻoihana mahiʻai organik i ʻoi aku ka nui o ka taxa (diamond) i hoʻohālikelike ʻia i ke koena, i kākoʻo i kona paʻakikī a me ka pilina.
Kiʻi 3. ʻO nā ʻupena fungal aʻa pili i ka ʻōnaehana mahiʻai
AUa ʻike ʻia ka pilina maikaʻi ʻole ma waena o ka ikaika o ka mahiʻai a me ka pilina pūnaewele fungal aʻa.Ua hōʻike ʻia ka nānā ʻana o ka ululāʻau i nā mea hoʻokele nui o ka ʻauhau pōhaku kiʻekiʻe: ka phosphorus lepo, ka nui nui, ka pH a me ka noho ʻana o ka mycorrhizal.
Kiʻi 4. ʻO ka ikaika o ka mahiʻai a me ka pilina pūnaewele ma waena o nā ʻōnaehana mahiʻai ʻekolu (A a me B);Ka nānā 'ana i ka nahelehele (C) a me ka pilina ma waena o ka ikaika o ka mahi'ai a me ka noho 'ana o AMF (D)
ʻenehana
Ke kaʻina amplicon lōʻihi piha
AE hele mai ana ka "Third Generation Sequencing" i ke kahua, ua lanakila nā palena o nā wahi i manaʻo ʻia a me nā pilikia i ka hui de novo.Ua hoʻonui maikaʻi ʻo Pacific Bioscience (PacBio) i ka heluhelu ʻana o nā kaʻina i nā ʻumi kilobases, e ʻae iā mākou e loaʻa i nā heluhelu holoʻokoʻa o 16s rRNA (1,000 bp-1,500 bp) i loko o ka bacteria a i ʻole 18S rRNA (1,500 bp-2,000 bp) a me ITS ʻāpana (400 bp-900 bp) i ka eukaryotics.ʻO ka ʻike ākea o ke kahua genetic i hoʻomaikaʻi nui i ka hoʻonā ʻana o nā ʻano kikoʻī a me nā genes hana.Ua hoʻoponopono ʻia ka pilikia lōʻihi e pili ana i ka pololei kumu e PacBio CCS hoʻoponopono ponoʻī, e hoʻopuka ana i nā heluhelu HIFI me ka 99% heluhelu pololei.
Hana ma ka hōʻike OTU
TʻO nā mea maikaʻi o ka heluhelu lōʻihi a me ka helu kiʻekiʻe, hiki ke hoʻonui nui ʻia ka pololei o ka annotation a loaʻa i kahi hoʻonā "species-level" i ka ʻike microbial.
Kuhikuhi
ʻO Banerjee, Samiran, et al."ʻO ka hoʻoikaika ʻana i ka mahiʻai e hōʻemi i ka paʻakikī o ka ʻoihana microbial a me ka nui o ka ʻauhau kīwī i nā aʻa."ʻO ka ISME Journal (2019).
ʻenehana a me nā mea koʻikoʻi manaʻo e kaʻana like i nā noi holomua hou loa o nā ʻenehana hoʻokaʻina kiʻekiʻe-throughput like ʻole ma nā ʻano ʻimi noiʻi like ʻole a me nā manaʻo maikaʻi loa i ka hoʻolālā hoʻokolohua a me ka ʻimi ʻikepili.
Ka manawa hoʻouna: Jan-08-2022