Luigi Ricciardi 1 , Rosa Mazzeo 2,*© , Angelo Raffaele Marcotrigiano 1 , Guglielmo Rainaldi 3 , Paolo Iovieno 4 , Vito Zonno 1 , Stefano Pavan 1© a me Concetta Lotti 2,*
- 1 'Oihana o ka lepo, mea kanu a me ka mea'ai 'epekema, Plant Genetics a me Breeding Unit University of Bari, Via Amendola 165/A, 70125 Bari, Italia; luigi.ricciardi@uniba.it (LR);angelo.marcotrigiano@uniba.it (ARM); vito.zonno@uniba.it (VZ); stefano.pavan@uniba.it (SP)
- 2 Keʻena ʻepekema o ka Mahiʻai, Meaʻai a me ke Kaiapuni, Ke Kulanui o Foggia, Via Napoli 25, 71122 Foggia, Italia
- 3 Keʻena o Biosciences, Biotechnologies and Biopharmaceuticals, University of Bari, Via Orabona 4, 70125 Bari, Italia; guglielmo.rainaldi@uniba.it
- 4 'Oihana o Energy Technologies, Bioenergy, Biorefinery and Green Chemistry Division, ENEA Trisaia Research Center, SS 106 Ionica, km 419+500, 75026 Rotondella (MT), Italia; paolo.iovieno@enea.it
* Kūlike: rosa.mazzeo@unifg.it (RM); concetta.lotti@unifg.it (CL)
Hōʻuluʻulu:
Onion (Allium cepa L.) ʻo ia ka lua o nā mea kanu mea kanu koʻikoʻi ma ka honua a mahalo nui ʻia no kāna mau pono olakino. ʻOiai ʻo kona koʻikoʻi koʻikoʻi a me kona waiwai ma ke ʻano he meaʻai hana, ua noiʻi maikaʻi ʻia ka onion e pili ana i kona ʻano genetic. Ma ʻaneʻi, ua nānā mākou i ka hoʻololi genetic i ka "Acquaviva red onion" (ARO), he landrace me ka moʻolelo o ke kenekulia o ka mahiʻai ʻana ma kekahi kūlanakauhale liʻiliʻi ma ka panalāʻau o Bari (Apulia, Hema o Italia). Ua hoʻohana ʻia kahi pūʻulu o nā māka microsatellite 11 no ka ʻimi ʻana i ka hoʻololi genetic i loko o kahi hōʻiliʻili germplasm i loaʻa i 13 mau heluna ARO a me ʻekolu mau ʻano kālepa maʻamau. Ua hōʻike ʻia nā ʻano loiloi o ka hoʻolālā genetic me nā ʻano parametric a me nā ʻano parametric ʻole i hōʻike ʻia ka ARO i kahi kolamu gene i wehewehe maikaʻi ʻia, ʻokoʻa loa mai nā ʻāina ʻo Tropea a me Montoro i kuhihewa pinepine ai. I mea e hāʻawi ai i ka wehewehe ʻana o nā ʻōpuʻu, i hoʻohana mau ʻia no ka ʻai hou ʻana, ua loiloi ʻia ka ʻiʻo paʻa paʻa a me ka pungency, e hōʻike ana i ka ʻono kiʻekiʻe o ka ARO e pili ana i nā ʻāina ʻelua i ʻōlelo ʻia ma luna. Ma ke ʻano holoʻokoʻa, pono ka haʻawina o kēia manawa no ka hoʻoikaika ʻana i ka ARO e hiki mai ana, hiki ke hoʻolaha ʻia ma o nā lepili maikaʻi e hiki ke kōkua i ka hoʻopaʻa ʻana i nā hoʻopunipuni kalepa a hoʻomaikaʻi i ka loaʻa kālā o nā mea liʻiliʻi.
Introduction
Aia ma kahi o 750 mau ʻano o ka Allium genus [1], a ma waena o ia ʻano aniani (Allium cepa L., 2n = 2x =16) ʻo ia kekahi o nā mea laha loa. He pōʻaiapuni ʻelua makahiki ko A. cepa a ma waho o ka hana hānau keiki. I kēia mau lā, ʻoi aku ka nui o ka ʻai ʻana i ka honua holoʻokoʻa (97.9 Mt) ʻo ia ka lua o nā mea kanu koʻikoʻi nui ma hope o ke kōmato [2]. Mai ka wā kahiko, ua hoʻohana ʻia nā ʻōpuʻu aniani ma ke ʻano he meaʻai a ma nā lāʻau lapaʻau. ʻOiaʻiʻo, ua hōʻike mua ka poʻe ʻAigupita kahiko i nā ʻano lapaʻau e pili ana i ka hoʻohana ʻana i ke kālika a me nā aniani i loko o kahi papyrus lāʻau lapaʻau o ka 1550 BC, ka Codex Ebers [3].
Hoʻohana ʻia kēia ʻano mea kanu maikaʻi a maikaʻi hoʻi i ka maka, hou, a i ʻole e like me ka huahana i hana ʻia, a hoʻohana ʻia e hoʻomaikaʻi i ka ʻono o nā kīʻaha he nui. Ua ʻōlelo kekahi mau noiʻi hou e hiki i ka ʻai ʻana ke hoʻemi i ka pilikia o nā maʻi cardiovascular [4,5], obesity [6], diabetes [7], a me nā ʻano maʻi like ʻole [8-10]. Hāʻawi pinepine ʻia nā waiwai olakino o ka onion i nā pae kiʻekiʻe o ʻelua papa o nā pūhui nutraceutical: flavonoids a me alk(en)yl cysteine sulphoxides (ACSOs). Aia ka papa mua i nā flavonols a me nā anthocyanins. ʻO Quercetin ka flavonol nui i ʻike ʻia, ʻike ʻia no kāna mau antioxidant ikaika a me nā waiwai anti-inflammatory i ka hoʻopaʻa ʻana i nā radical manuahi a me ka hoʻololi ʻana i nā ion metala. [11]; ʻoiai hāʻawi nā anthocyanins i ka ʻulaʻula/ʻulaʻula i kekahi mau ʻano aniani. No nā ACSO, ʻo ka nui loa ka isoalliin [(+)-trans-S-1-propenyl-L-cysteine sulfoxide] [12], he waika amino sulfur non-volatile a non-proteinogenic i mālama ʻia i loko o nā keena, nona ke kuleana no ka ʻala ʻala a me ka ʻono o nā aniani. [13]. Ma hope o ka hoʻohaunaele ʻana o ka ʻiʻo, hoʻokaʻawale ʻia ka isoalliin e ka enzyme alliinase e hana i kahi pūʻulu o nā mea hoʻoheheʻe (pyruvate, ammonia, thiosulphonates a me propanethial S-oxide) e hoʻoulu ai i ka haehae ʻana a me ka honi maikaʻi ʻole. [14]. ʻIke pinepine ʻia ka pungency onioni e like me ka nui, no ka gram o ke kaumaha hou, o ka waika pyruvic i hana ʻia e ka hydrolysis. [15,16].
Ma nā ʻāina o ke kai Mediterranean, ua manaʻo ʻia ʻo ia kekahi o nā kikowaena ʻokoʻa lua o A. cepa [17,18], Hōʻike nā ʻōpuʻu aniani i ka ʻokoʻa ākea o ke ʻano, ka nui, ka waihoʻoluʻu, ka mea maloʻo, a me ka pungency [19-ʻekahi]. Eia kekahi, hiki i ka fertilization-based fertilization, agronomic practices, type of soil, climatic condition, and the genotype of cultivars or landraces ke hoʻoikaika i ka maikaʻi o ka puʻupuʻu ma o ka hāʻawi ʻana i nā waiwai organoleptic a me ka meaʻai. [23-ʻekahi]. Ma Italia, ʻoiai ka loaʻa ʻana o ka germplasm onion ākea, he liʻiliʻi wale nō nā ʻano aniani i hoʻopaʻa pinepine ʻia i nā noiʻi ʻepekema a hōʻike pono ʻia. [28,29].
He mea koʻikoʻi ke ʻano genetic a me ka phenotypic o ka agro-biodiversity e hōʻoiaʻiʻo i ka mālama pono ʻana i nā kumu waiwai mea kanu a paipai i ka hoʻohana ʻana i nā genotypes kikoʻī i ke kaulahao waiwai. [30-ʻekahi]. Ua koho pinepine ʻia nā hōʻailona hana hou maʻalahi (SSR) no ka palapala ʻāina [33-ʻekahi], DNA manamana lima a me ka hoʻokae ʻana [36-ʻekahi], a me ka manaʻo hilinaʻi o ka ʻokoʻa genetic i loko a ma waena o nā ʻāina [39-ʻekahi], no ka mea, he wahi kiko'ī lākou, multi-allelic, codominantly hoʻoilina, hiki ke hana hou ʻia, a kūpono no ka genotyping automated.
Ma ka haʻawina o kēia manawa, ua kālele mākou i ko mākou manaʻo i kahi ʻāina kuʻuna Apulian, ʻo "Acquaviva red onion" (ARO), i mahi ʻia e like me nā ʻano mahiʻai organik ma kahi wahi liʻiliʻi o ke kūlanakauhale ʻo Acquaviva delle Fonti, ma ka ʻāina ʻo Bari. (Apulia, ʻIlia Hema). ʻO nā ʻōpuʻu o kēia landrace he nui a palahalaha a ʻulaʻula a hoʻohana nui ʻia i nā ʻano ʻai kūloko. ʻOiai ua loaʻa i ka ARO ka hōʻailona maikaʻi ʻo "Slow Food Presidium", hiki ke hoʻonui a pale ʻia kāna hana ʻana e nā hōʻailona maikaʻi o ka European Union e like me ka protected geographical indication (PGI) a me ka protected designation of origin (POD), no ka mea e kōkua paha kēia i ka palena hoʻopunipuni kālepa a hoʻomaikaʻi i ka loaʻa kālā o nā mea liʻiliʻi. Ma ʻaneʻi, ua hoʻohana ʻia nā māka molekala SSR ma ke ʻano he mea hana ikaika e loiloi i ka hoʻololi ʻana o ka genetic ma waena o nā heluna ARO a no ka hoʻokaʻawale ʻana i kēia ʻāina mai nā ʻāina ʻulaʻula ʻulaʻula ʻelua o ka Hema. Eia kekahi, ua manaʻo mākou i ka pungency a me ka ʻike paʻa paʻa i mea e loiloi ai i ka ʻono ARO e pili ana i ka noi mākeke.
Results
Hoʻokumu ʻia ʻo Acquaviva Red Onion Germplasm Collection a me Morphological Characterization
Ua hoʻohana ʻia nā hua o 13 heluna kanaka o ka ʻāina ARO, i hāʻawi ʻia e ka poʻe mahiʻai ma ke ʻano o ka papahana BiodiverSO Apulia Region e hoʻokumu i kahi hōʻiliʻili ARO germplasm.
Ua hōʻiliʻili ʻia nā mea wehewehe moʻokalaleo, pili i ka puʻupuʻu, ka ʻili, a me ka ʻiʻo ma luna o ka germplasm ARO a ma luna o ʻekolu landrace onion, ʻelua no ka "Tropea red onion" (TRO) landrace a hoʻokahi no ka "Montoro copper onion" (MCO) landrace (Figure 1). ʻO nā puʻupuʻu ARO a pau he palahalaha a ua ʻike ʻia e ka ʻili waho ʻulaʻula a me ka ʻiʻo me nā ʻano ʻulaʻula like ʻole. ʻO ka ʻokoʻa, ʻulaʻula piha ka ʻiʻo o nā puʻupuʻu TRO, a ʻo ka ʻiʻo o nā puʻupuʻu MCO ua ʻulaʻula maikaʻi ʻia (Pable S1). Ua ʻae ʻia ka loiloi biochemical e loiloi i ka ʻike paʻa paʻa a me ka pungency. E like me ka hoike ma ka Papa 1, ʻo ka nui o ka nui o ka maʻiʻo paʻa hiki ke hoʻoheheʻe ʻia o nā puʻupuʻu i nā heluna ARO he 7.60, a mai ka 6.00 (ARO12) a i ka 9.50° Brix (ARO11 a me ARO13). Ua ʻoi aku kēia waiwai ma mua o ka mea i manaʻo ʻia no nā ʻāina TRO a me MCO (4.25 a me 6.00° Brix, kēlā me kēia).
1 Pūnaewele. ʻIke ʻia nā maʻiʻo paʻa paʻa a me nā waiwai Pungency ma "Acquaviva Red Onion" (ARO), "Tropea Red Onion" (TRO), a me "Montoro Copper Onion" (MCO) Populations *.
KANAWAI | Maʻiʻo Paʻa Paʻa (Brix) | Pungency (pmolg-1 FW) | ||
Mean | CV y (%) | Mean | CV y (%) | |
ARO1 | 6.25 D * | 5.65 | 5.84 ab * | 23.78 |
ARO2 | 7.25 DC | 4.87 | 6.51 i | 22.98 |
ARO3 | 7.50 BCD | 9.42 | 5.28ab | 22.88 |
ARO4 | 7.50 BCD | 0.00 | 6.97 i | 3.74 |
ARO 5 | 7.50 BCD | 0.00 | 6.80 i | 9.68 |
ARO6 | 6.25 D | 5.65 | 4.51ab | 39.18 |
ARO7 | 7.25 DC | 4.87 | 5.25ab | 15.44 |
ARO8 | 9.00 AB | 0.00 | 7.04 i | 3.49 |
ARO9 | 8.25 ABC | 4.28 | 6.84 i | 0.15 |
ARO10 | 7.00 DC | 0.00 | 5.94ab | 6.57 |
ARO11 | 9.50 A | 7.44 | 5.54ab | 16.43 |
ARO12 | 6.00 D | 0.00 | 4.91ab | 9.70 |
ARO13 | 9.50 A | 7.44 | 6.63 i | 24.93 |
MCO | 6.00 D | 0.00 | 4.18ab | 2.66 |
TRO1 | 4.25 E | 8.31 | 2.80 b | 2.10 |
TRO2 | 4.25 E | 8.31 | 4.28ab | 4.79 |
* ʻAʻole ʻokoʻa ʻokoʻa nā huaʻōlelo me nā leka like ma ka ʻaoʻao nui a i ʻole ka liʻiliʻi ma 0.01P a i ʻole 0.05P (SNK's Test). y Coefficient o ka hoʻololi.
ʻO ka nui o ka waiwai o ka ARO pungency, i helu ʻia e ka pyruvic acid content, he 6.00, mai ka 4.51 pmol g.-1 FW (ARO6) i ka 7.04 (ARO8). Ua ʻoi aku ka kiʻekiʻe o kēia waiwai ma mua o ka mea i manaʻo ʻia ma nā ʻāina TRO a me MCO (3.54 pmol g-1 FW a me 4.18 pmol g-1 FW, kēlā me kēia).
SSR Polymorphism a me Genetic Relationships between Accessions
Ma ke aʻo ʻana i kēia manawa, 11 o 37 i hoʻāʻo ʻia nā hui kumu SSR i hoʻolako i nā polymorphisms locus hoʻokahi, ʻo ia hoʻi, e hāʻawi ana i ka nui o nā huahana amplification ʻelua i hoʻokahi kanaka. ʻO ka holoʻokoʻa, ua ʻike ʻia nā 55 alleles i 320 mau kānaka me ka nui o nā alleles ma kēlā me kēia locus mai 2 (ACM147 a me ACM 504) a i 11 (ACM132) a me kahi waiwai o 5 alleles (Papa. 2). Ma ka heluna kanaka pākahi, ʻo ka helu o nā alleles (Na) mai 1.94 (ACM147 a me ACM504) a i 5.38 (ACM132), aʻo ka helu kūpono o nā alleles (Ne) mai 1.41 (ACM152) a i 2.82 (ACM449). Kūlike ʻole ma waena o nā koina Na a me Ne ma muli o ka loaʻa ʻana o nā alleles me ke alapine haʻahaʻa i loko o nā heluna kanaka a me ka predominance o nā alleles wale nō. Ua hōʻike ʻia ka waiwai heterozygosity (Ho) kiʻekiʻe loa no ACM138 a me ACM449 (0.62), aʻo ka mea haʻahaʻa i pili me ACM152 (0.25). ʻO ka heterozygosity i manaʻo ʻia (He), e pili ana i ka manaʻo manaʻo i ka heluna panmictic, mai 0.37 (ACM504) a i 0.61 (ACM132, ACM138, a me ACM449). Ua hōʻike ʻo Wright's fixation index (Fis) i nā waiwai kokoke i ka ʻole (0.05 awelika) no nā māka āpau, e hōʻike ana i nā waiwai like ma waena o nā pae heterozygosity i ʻike ʻia a i manaʻo ʻia, e like me ka mea i manaʻo ʻia no kahi ʻano outcrossing. Ua manaʻo ʻia ka maikaʻi o kēlā me kēia SSR marker ma ka genetic fingerprinting e ka polymorphic information content (PIC) index, me ka waiwai nui o 0.48 a mai ka 0.33 (ACM504) a i ka 0.67 (ACM132). ʻO kekahi papa kuhikuhi kūpono, ʻo ka Shannon's Information Index (I) i hōʻike i ka waiwai mean o 0.84, a ua manaʻo ʻia nā waiwai mai ka 0.45 (ACM152) a i ka 1.20 (ACM132).
2 Pūnaewele. ʻO nā hiʻohiʻona Polymorphism o nā 11 SSR Markers i hoʻohana ʻia no ka hoʻohālikelike ʻana i ka ʻokoʻa Genetic ma ARO, TRO, a me MCO heluna kanaka. Ka heluna nui o nā Alleles (Na), ka nui o ka Band, a me ka Polymorphic Information Content (PIC) Index E nānā i ka huina o 320 mau kanaka i Genotyped i kēia Haʻawina. Ka helu o nā Alleles (Na), ka helu o nā Alleles Effective (Ne), Heterozygosity i ʻike ʻia (Ho), Heterozygosity i manaʻo ʻia (He), Fixation Index (Fis), a me Shannon's Information Index (I) e pili ana i ka Mean Values Calculated from 16 Populations, kela mea keia mea i hakuia e 20 kanaka.
Locus. | Huina Na | Nui (bp) | pic | Mean | |||||
Na | Ne | Ho | He | I | Fis | ||||
ACM91 | 4 | 189-205 | 0.40 | 2.63 | 1.72 | 0.38 | 0.39 | 0.66 | 0.04 |
ACM101 | 4 | 229-241 | 0.52 | 2.94 | 2.37 | 0.53 | 0.56 | 0.92 | 0.06 |
ACM132 | 11 | 186-248 | 0.67 | 5.38 | 2.78 | 0.55 | 0.61 | 1.20 | 0.09 |
ACM138 | 5 | 242-272 | 0.66 | 3.69 | 2.82 | 0.62 | 0.61 | 1.09 | -0.02 |
ACM147 | 2 | 264-266 | 0.37 | 1.94 | 1.83 | 0.44 | 0.44 | 0.62 | -0.01 |
ACM152 | 4 | 228-244 | 0.25 | 2.38 | 1.41 | 0.25 | 0.27 | 0.45 | 0.07 |
ACM235 | 4 | 286-298 | 0.41 | 2.81 | 1.77 | 0.44 | 0.41 | 0.72 | -0.06 |
ACM446 | 6 | 108-120 | 0.56 | 3.50 | 2.48 | 0.49 | 0.58 | 1.01 | 0.16 |
ACM449 | 8 | 120-140 | 0.66 | 4.88 | 2.82 | 0.62 | 0.61 | 1.18 | -0.03 |
ACM463 | 5 | 202-210 | 0.47 | 3.38 | 1.95 | 0.46 | 0.48 | 0.83 | 0.05 |
ACM504 | 2 | 188-192 | 0.33 | 1.94 | 1.64 | 0.30 | 0.37 | 0.54 | 0.20 |
Mean | 5 | 0.48 | 3.22 | 2.15 | 0.46 | 0.48 | 0.84 | 0.05 |
Ma waena o nā heluna kanaka, ʻo ARO3, ARO6, ARO8, ARO10, TRO1, a me MCO i hōʻike i ke kiʻekiʻe o ka hoʻololi genetic (Ho > 0.5), ʻoiai ʻo ka ʻokoʻa haʻahaʻa i ʻike ʻia i ka heluna ARO7 (Ho = 0.27) (Paʻa S2). Ma keʻano holoʻokoʻa, ua hōʻike ʻia nā komo a pau iā Fis nā waiwai kokoke i ka ʻole (Fis kumu waiwai = 0.054), e like me ka mea i manaʻo ʻia ma lalo o nā kūlana hoʻohālikelike.
ʻIkepili o ka ʻokoʻa Molecular a me ke ʻano Genetic Structure
ʻO ka hoʻokaʻawale hierarchical o ka hoʻololi genetic i waena a i loko o ka heluna kanaka i helu ʻia e AMOVA. Ua hōʻike ʻia nā hopena i kahi hapa nui o ka hoʻololi genetic i loko o nā heluna (87%). ʻO ka ʻokoʻa ma waena o nā heluna, 13%, he mea nui loa (P <0.001) (Papa 3). ʻO nā waiwai pālua o ka pānaʻi Fpt, he mea like me ka papa kuhikuhi hoʻoponopono Fst o Wright, mai ka 0.002 (ARO2/ARO10) a i ka 0.468 (ARO7/TRO2), he mea nui (P <0.05), koe wale no ʻeiwa mau hoʻohālikelike hoʻohālikelike (S3 Pākuʻi Pākuʻi).
3 Pūnaewele. Ka Ikepili o ka Molecular Variance o 320 Genotypes mai 16 Populapula o allium cepa L.
kahi | df | Huina o nā Kaha | Hoʻohālikelike ʻokoʻa | ʻokoʻa (%) | Fpt | P |
Ma waena o ka lehulehu | 15 | 458.63 | 1.16 | 13% | ||
Iloko o na kanaka | 304 | 2272.99 | 7.50 | 87% | 0.134 | 0.001 |
Ka huina | 319 | 2731.62 | 8.66 |
Ke noiʻi ʻana i ke ʻano genetic i loko o ka A. cepa ʻO ka hōʻiliʻili genotyped i kēia haʻawina i hana ʻia ma ke ʻano o ka hoʻohui ʻana i nā kumu hoʻohālike i hoʻokō ʻia i ka polokalamu STRUCTURE. Ua manaʻo ke ala Evanno AK i ka hoʻokaʻawale ʻana i ʻelua mau puʻupuʻu (K = 2) ma ke ʻano he ʻike nui loa no kā mākou. kuapo,me the aʻe kiʻekiʻe loa peak ma K = 5 (ʻo Rgure hoʻohui S1). A no K = 2, ahpopulations wmua hokiigned i ona me kaf nā pūʻulu ʻelua me he huinahelu rnernbertoip (q) > 0.7. E like me shown i loko Hōʻike 2a, ʻo ka hui mua (i kapa ʻia ʻo S1) i hui pū ʻia me MCO a me nā heluna ARO a pau, ʻoiai ua hui pū ka pūʻulu S2 i nā heluna TRO ʻelua. Ma K = 5, e hāʻawi ana i kahi wehewehe hohonu o ka waihona (Figure 2b), 75% o nā komo ʻana i hāʻawi ʻia i hoʻokahi o nā pūʻulu ʻelima. Ua hōʻoia ʻia ka hoʻokaʻawale ʻana ma waena o ARO (S1) a me TRO (S2), ʻoiai ua hoʻohui ʻia kekahi mau heluna ARO (q <0.7) a i hui pū ʻia paha i loko o nā hui hou ʻelua S3 a me S4 (ARO7 a me ARO12, kēlā me kēia). ʻO ka mea e mahalo ai, ua hoʻokumu ʻia ka ʻano pāʻoihana MCO i pūʻulu ʻokoʻa (S5) i hoʻokaʻawale ʻia mai ke aniani ʻulaʻula Apulian.
Nā Pili Genetic ma waena o nā Lehulehu
Ua ʻae ʻia ka polymorphism SSR e huki i kahi dendrogram o ka ʻokoʻa genetic a hōʻike ʻia nā hopena o ka loiloi phylogenetic ma ke Kiʻi. 3a. Ma ʻaneʻi, ua māhele ʻia ka ʻohi germplasm i ʻelima mau pūʻulu i kākoʻo ikaika ʻia e nā waiwai bootstrap. Ua hoʻokaʻawale koke ʻia nā heluna ARO7 a me ARO12 mai nā heluna i koe a ua hana ʻia ʻelua mau pūʻulu ʻokoʻa. ʻO ke kolu o ka puʻupuʻu i loaʻa nā heluna kanaka kalepa ʻelua o TRO, ʻoiai ka hā o ka node i hoʻokaʻawale iā MCO mai ʻumikūmākahi mau heluna ARO. Ua noiʻi hou ʻia ka pilina genetic i waena o ka heluna kanaka ma o ke kumu hoʻohālikelike kumu (PCoA) (Figure 3b). E like me ka mea i hōʻike mua ʻia, ua hui pū ʻia nā heluna ARO, koe wale nō ʻo ARO12 a me ARO7, i ʻike ʻia ma nā kūlana kaʻawale i ka pā PCoA. Ua hoʻopuehu ʻia nā TRO ʻelua a me ka heluna MCO ma ka ʻaoʻao ʻākau ʻākau o ka pā.
Hōʻike 3. ʻO ka ʻokoʻa genetic ma waena o 16 A. cepa ka heluna kanaka i hōʻike ʻia i kēia haʻawina, e pili ana i kā lākou SSR profile. (a) UPGMA dendrogram o ka mamao genetic. Hōʻike ʻia nā waiwai kākoʻo Bootstrap> 50 ma luna o nā nodes pili; (b) ka hoʻolālikelike ʻāpana kumu (PCoA). Ua hoʻopili piha ʻia ka puʻupuʻu i ka ʻulaʻula me ka hui i hana ʻia e ka phylogenetic analysis a ua hoʻokumu ʻia e 11 mau komo ARO.
kūkākūkā
I loko o ka nui o ka agro-biodiversity i kanu ʻia ma ka Hema Italia, ʻo nā ʻāina ʻonika e hōʻike ana i nā huahana niche e pono e mālama ʻia mai ka hopena o ka hoʻoneʻe genetic a me ka hoʻoweliweli o ka hoʻololi ʻana e nā cultivars hou. Ma ke ʻano o ka papahana ʻāpana ʻo BiodiverSO, i manaʻo ʻia e hōʻiliʻili, hōʻike, hoʻolaha, a me ka mālama ʻana i nā kumuwaiwai genetic o ka ʻāina ʻo Apulia i pili ikaika i ka hoʻoilina kūloko, ua hoʻokumu mākou i kahi hōʻiliʻili hua o 13 mau kānaka o ka ʻāina ARO. Ua hōʻike mākou i ka loiloi mua o ka ʻokoʻa ARO e pili ana i nā polymorphism DNA a me ʻelua mau ʻāpana biochemical, soluble solid a pyruvic acid contents, pili i nā ʻano ʻono a me ka mea nui no ka ʻae ʻana i nā huahana hou ʻole. Eia kekahi, ua hoʻohālikelike ʻia nā ʻikepili ma ka ʻāina ʻo ARO me nā mea i hōʻiliʻili ʻia ma nā ʻāina ʻonika ʻelua ʻē aʻe i kuhi hewa ʻia.
Ua hōʻike ʻia nā loiloi biochemical i ka ʻono o nā heluna ARO he 13, pili i ka mea paʻa paʻa soluble kiʻekiʻe a me ka pungency waena, e like me nā alakaʻi alakaʻi ʻoihana ʻona. [31]. Ua ʻoi aku ka ʻono o nā puʻupuʻu ARO ma mua o nā ʻāina TRO a me MCO, a ua hōʻike ʻia i kahi pungency ʻoi aʻe. Eia nō naʻe, ʻo ka ʻono i loko o nā aniani ma muli o ke kaulike ma waena o ke kō a me ka pungency, no laila hiki ke hoʻohana ʻia kēia hiʻohiʻona e kākoʻo i ke koho ʻana i nā genotypes o ka waiwai, e hana mau ʻia e nā mahiʻai ma muli o ka morphology.
Ua hōʻoia ʻia nā māka SSR he mea hana pono e hoʻokaʻawale i nā genotypes, ʻoiai e hōʻiliʻili ʻia i loko o kahi wahi ulu haiki e like me ke kaona ʻo Acquaviva delle Fonti. Ua hōʻike ʻia nā māka i koho ʻia i ka helu ʻoi aku o nā allele ma mua o nā māka i hōʻike mua ʻia e [43] a [44], akā, haʻahaʻa ma mua o nā māka i hōʻike ʻia e [45]. Eia kekahi, 50% o kā mākou pūʻulu o nā hōʻailona hōʻike i nā helu helu PIC i ʻoi aku ma mua o 0.5, e hōʻike ana he kūpono ia e hoʻokaʻawale i nā heluna o ka hōʻiliʻili, e like me ka manaʻo [46]. Ua hōʻike ʻia ka loiloi o ka ʻokoʻa i loko o ka heluna kanaka i nā waiwai like ma waena o Ho a me He, a ua haʻahaʻa ka Fis waiwai. He kulike keia me ke ano o ka hele ana i waho A. cepa, ka mea e pilikia nui ana i ke kaumaha inbreeding [47]. ʻO ka Fis ʻO ka waiwai i helu ʻia ma nā heluna ʻonika i manaʻo ʻia ma kēia haʻawina (0.054) ʻoi aku ka haʻahaʻa ma mua o ka mea i hōʻike mua ʻia e [45] (0.22) a aneane like me ka mea i loaa e [31] (0.08) a me ka [48] (0.00) nāna i loiloi i ka ʻokoʻa ʻokoʻa i nā ʻāina ʻonika mai ke komohana ʻākau o Sepania a me Niger, kēlā me kēia. ʻO nā pae koʻikoʻi o ka heterozygosity i nā heluna ARO e hoʻoikaika i ka manaʻo e hōʻike ana ʻo Apulia i kahi kikowaena like ʻole no nā ʻano horticultural he nui. [32, 42, 49-ʻekahi].
Ua hōʻike ʻo AMOVA i ka nui o nā ʻano molekala i loko o ka hōʻiliʻili genotyped i kēia haʻawina aia i loko o nā heluna. Eia nō naʻe, ʻo ka ʻokoʻa genetic koʻikoʻi ma waena o nā heluna (FPT nā waiwai) i hōʻike i ka hiki ʻana o ka stratification genetic. ʻO ka ʻoiaʻiʻo, ʻoiai ua hōʻike kā mākou mau hopena i ka loaʻa ʻana o ka hoʻohālikelike genetic i ka hapa nui o nā heluna ARO, e hana ana i kahi puʻupuʻu i wehewehe maikaʻi ʻia, ua hōʻike ʻia nā heluna ARO7 a me ARO12 i kahi moʻo kikoʻī kikoʻī. Ma muli o ke kumu ʻokoʻa o nā hua i hoʻohana ʻia e nā mahiʻai ʻelua i hōʻiliʻili ʻia ai ka heluna kanaka. Eia kekahi, ma muli o nā hopena i loaʻa, hiki ke noʻonoʻo ʻia ka ʻāina ʻāina ʻo ARO i ka ʻokoʻa ma ka pae genetic mai nā ʻāina TRO a me MCO. Ma kahi noiʻi hou, [29] ua loiloi i ka ʻokoʻa ʻokoʻa o nā ʻāina ʻonika ʻItalia me "Acquaviva," "Tropea," a me "Montoro." ʻOiai ua hoʻohana nā mea kākau i nā māka SNP e loiloi i ka ʻokoʻa genetic o kahi hōʻiliʻili aniani ākea, ʻaʻole hiki i ka genotyping ke hoʻokaʻawale i ka "Acquaviva" mai "Tropea" a me "Montoro". Malia paha, ʻo kēia ʻokoʻa ma muli o ka haʻahaʻa haʻahaʻa PIC waiwai i loaʻa (0.292), e hōʻike ana i kahi ʻike ākea haʻahaʻa o ka loci ma lalo o ka nānā ʻana e like me ka ʻōlelo a [29]. Eia kekahi, i mea e noiʻi ai i ke ʻano o ka sub-structure i loko o kā lākou hui Italia, ʻoi aku ka maikaʻi o ka nānā ʻana i nā genotypes Italia ma kahi kaʻawale mai ke koena o ka hōʻiliʻili. Malia paha ua ʻae ʻia ke ʻike ʻana i ke ʻano o ka ʻokoʻa genetic i pili i ka stratification ʻāina a i ʻole nā hiʻohiʻona ma lalo o ke koho empiric.
I ka hopena, hōʻike ka haʻawina i kēia manawa i kahi hōʻike piha e pili ana i kahi ʻāina aniani e pili ana i ka hoʻoilina moʻomeheu kūloko a me ka waiwai waiwai no ka poʻe mahiʻai. Hōʻike kā mākou mau hualoaʻa, me nā ʻokoʻa liʻiliʻi, ʻike ʻia ʻo ARO e kahi pūnāwai gene i wehewehe maikaʻi ʻia, pono e mālama ʻia mai ka hopena o ka erosion genetic. No laila, he mea koʻikoʻi ka hoʻokumu ʻana i kahi hōʻiliʻili ʻelele o kēia kumu waiwai o ka ʻokoʻa genetic. ʻO ka mea hope loa, hiki ke hoʻohana ʻia ke ʻano genetic a me ka phenotypic o ARO e kiʻi i nā hōʻailona maikaʻi mai ka European Union.
Nā Pono a me nā Una
Hōʻiliʻili ʻo Germplasm, Mea Mea kanu, a me ka ʻili DNA
Ua loaʻa kahi pūʻulu o 13 mau kānaka o ka ʻāina ʻāina ʻo ARO i loko o ke ʻano o kahi papahana Apulia Region (BiodiverSO: https://www.biodiversitapuglia.it/), ma o ke kaʻina o nā mikiona i hana ʻia ma "Acquaviva delle Fonti", he kūlanakauhale liʻiliʻi ʻo Apulian ma ka moku o Bari, Italia. Ua palapala ʻia nā pae ohi o kēlā me kēia komo ʻana ma o Geographic Information System (GIS) a hōʻike ʻia ma ka Papa 4. Eia kekahi, ʻelua heluna kanaka mai ka TRO landrace a me hoʻokahi heluna kanaka mai ka MCO landrace i hoʻokomo ʻia i loko o ka haʻawina o kēia manawa a hoʻohana ʻia i mau kuhikuhi. Hoʻoulu ʻia nā mea kanu āpau i nā kūlana kaiapuni like ma ka māla hoʻokolohua "P Martucci" o ke Kulanui o Bari (41° 1'22.08″ N, 16°54'25.95″ E), ma lalo o ka hale hoʻomalu e pale aku i ka pollination ma waena ka heluna kanaka a me ka hōʻoiaʻiʻo ʻana i ka pollination intra-population ma o nā lele lele (Lucilia Kaisara). Ua ʻike ʻia ka heluna kanaka he 16 no nā hiʻohiʻona pili i ka nui a me ke ʻano o ka ʻili a me ka ʻiʻo (Papa S1). Eia kekahi, ua hoʻohana ʻia ka hoʻohana ʻana i ka refractometer paʻa i ka lima a ua ana ʻia ka pungency i loko o nā ʻāpana wai aniani e hoʻohui ana i ka 2,4-dinitrophenyl hydrazine (0.125%). v/v i 2N o HCl) a me ka loiloi ʻana i ka absorbance ma 420 nm, e like me ka mea i hōʻike ʻia e [31]. Ua hoʻokō ʻia ka hoʻāʻo ʻana o Duncan a me ka hoʻāʻo SNK e hoʻoholo ai i ka loaʻa ʻana o nā ʻokoʻa nui.
4 Pūnaewele. Ka papa inoa o nā poʻe i hōʻiliʻili ʻia a hoʻopaʻa ʻia i loko o kēia haʻawina. No kēlā me kēia heluna kanaka, hōʻike ʻia ke Code Identification, Local Name, GPS Coordinate, a me Gene Bank Preserving the Seeds.
Code | inoa | Nā Koina GPS | Gene Bank y |
ARO1 | Cipolla rossa ma Acquaviva | 40°54’21.708″ N 16°49’1.631” E | Di.SSPA |
ARO2 | Cipolla rossa ma Acquaviva | 40°53’14.28″ N 16°48’56.879” E | Di.SSPA |
ARO3 | Cipolla rossa ma Acquaviva | 40°54’11.304″ N 16°49’13.079” E | Di.SSPA |
ARO4 | Cipolla rossa ma Acquaviva | 40°54’3.348″ N 16°40’27.011” E | Di.SSPA |
ARO5 | Cipolla rossa ma Acquaviva | 40°51’59.76″ N 16°53’0.527” E | Di.SSPA |
ARO6 | Cipolla rossa ma Acquaviva | 40°52’48.72″ N 16°49’43.247” E | Di.SSPA |
ARO7 | Cipolla rossa ma Acquaviva | 40°53’13.47″ N 16°50’23.783” E | Di.SSPA |
ARO8 | Cipolla rossa ma Acquaviva | 40°53’18.816″ N 16°49’33.888” E | Di.SSPA |
ARO9 | Cipolla rossa ma Acquaviva | 40°54'51.372″ N 16°49'3.504" E | Di.SSPA |
ARO10 | Cipolla rossa ma Acquaviva | 40°54’1.188″ N 16°49’24.311” E | Di.SSPA |
ARO11 | Cipolla rossa ma Acquaviva | 40°52'49.8″ N 16°49'48.575" E | Di.SSPA |
ARO12 | Cipolla rossa ma Acquaviva | 40°52’38.892″ N 16°49’28.379” E | Di.SSPA |
ARO13 | Cipolla rossa ma Acquaviva | 40°53’21.768″ N 16°49’29.711” E | Di.SSPA |
TRO1 | Cipolla rossa lunga ma Tropea | - | Di.SSPA |
TRO2 | Cipolla rossa tonda di Tropea | - | Di.SSPA |
MCO | Cipolla ramata di Montoro | - | Di.SSPA |
y Di.SSPA, Keʻena o ka lepo, mea kanu a me ka ʻepekema meaʻai, Kulanui o Bari. |
ʻO nā lau lau o 20 genotypes no kēlā me kēia heluna kanaka i hōʻike ʻia a mālama ʻia ma -80 °C a hiki i ka hoʻohana ʻana. No nā ʻano mea waiwai polysaccharide, e like me A. cepa, ʻO nā hana mua e wehe i ka polysaccharide he mea nui ia e loaʻa ai ka DNA maikaʻi, no laila ua hana ʻia ka holoi mua ʻana i ka STE buffer (0.25 M sucrose, 0.03 M Tris, 0.05 M EDTA) e like me ka wehewehe ʻana. [52]. Ua unuhi ʻia ka huina DNA ma muli o ke ʻano CTAB [53] a hope loa ua nānā ʻia no ka maikaʻi a me ka noʻonoʻo ʻana e Nano Drop 2000 UV-vis spectrophotometer (ThermoScientific, Waltham, MA, USA) a me 0.8% agarose gel electrophoresis.
Nānā SSR
16 EST-SSR hui mua i haku ʻia e [54] a ua hoʻāʻo mua ʻia ma nā haʻawina ʻano like ʻole e [43] a [44] a me 21 genomic SSR [45-ʻekahi] ua nānā ʻia no ka loiloi ʻana i ko lākou kūpono (Supplementary Table S4). Hoʻohana ʻia ka genotyping me ka hoʻohana ʻana i ke ʻano hana hōʻailona fluorescent e hoʻohui ʻia ka huelo M13 i kēlā me kēia SSR mua. [56]. Ua hoʻomākaukau ʻia nā huila PCR i 20 gL hoʻololi i loaʻa: 50 ng o ka huina DNA, 0.2 mM o ka hui dNTP, 1X o ka PCR reaction buffer, 0.8 U o DreamTaq DNA polymerase (Thermo Scientific, Waltham, MA, USA), 0.16 gM o ka reverse primer. , 0.032 gM o ka mua mua i hoʻonui ʻia me ke kaʻina M13 (5′-TGTAAAACGACGGCCAGT-3′), a me 0.08 gM o ka M13 kumu mua i hoʻopaʻa inoa ʻia me FAM a i ʻole NED nā mea hoʻonani fluorescent (Sigma-Aldrich, St. Louis, MO, USA). Ua hoʻokō ʻia nā hopena PCR i ka thermocycler SimpliAmp (Applied Biosystems, CA, USA) me nā kūlana aʻe no ka hapa nui o nā hui mua: 94 °C no 5 min, 40 cycles ma 94 °C no 30 s, 58 °C no 45 s a me 72 °C no 45 s a me ka elongation hope ma 72 °C no 5 min. E like me ACM446 a me ACM449, ua hoʻohana ʻia kahi PCR touchdown me ka annealing o 60 °C a i 55 °C ma luna o 10 cycles, 30 cycles ma 55 °C, a ukali ʻia e ka hoʻonui hope o 5 min ma 72 °C. Hoʻokomo ʻia nā huahana PCR i loko o kahi pā 96-well a hui pū ʻia me 14 gL o Hi-Di Formamide (Life Technologies, Carlsbad, CA, USA) a me 0.5 gL GeneScan 500 ROX Size Standard (Life Technologies, Carlsbad, CA, USA). Ua hoʻoholo ʻia nā Amplicons e ka ABI PRISM 3100 Avant Genetic Analyzer (Life Technologies, Carlsbad, CA, USA) capillary sequencing machine, kahi i helu ʻia ai nā alleles ma ke ʻano he co-dominant a hāʻawi ʻia e ka hoʻohana ʻana i ka GeneMapper Software Version 3.7.
ʻO nā polokalamu GenAlEx 6.5 [57] a me Cervus 3.0.7 [58] ua hoʻohana ʻia no ka helu ʻana i ka helu o nā alleles (Na), ka helu o nā alleles kūpono (Ne), ʻike ʻia ka heterozygosity (Ho), heterozygosity i manaʻo ʻia (He), polymorphic information content (PIC), Shannon's information index (I), a me fixation index (Fis). ) no kēlā me kēia wahi SSR.
Ka Loiloi o ka Genetic Diversity
Ua loiloi ʻia e GenAlEx 6.5 ka hoʻokaʻawale hierarchical o ka hoʻololi genetic i waena a i loko o ka heluna ʻonika. [57] ma o ke kālailai ʻana o ka molecular variance (AMOVA) me 999 bootstrapping e hoʻāʻo ai no ke koʻikoʻi. Eia kekahi, ua hoʻohana ʻia ka polokalamu GenAlEx 6.5 e koho i ka ʻokoʻa i loko o kēlā me kēia heluna kanaka ma o ka helu ʻana i ka awelika o Ho, He, a me Fis ma luna o nā wahi SSR āpau.
Ua ʻike ʻia ka hoʻolālā heluna kanaka e ka Bayesian model-based clustering algorithm i hoʻokō ʻia ma ka polokalamu STRUCTURE v.2.3.4. [59]. Ua holo ka ʻikepili me ka nui o nā puʻupuʻu hypothetical (K), mai ka 1 a hiki i ka 10, e hoʻonohonoho ana i ʻumi holo kūʻokoʻa no kēlā me kēia K waiwai. No kēlā me kēia holo, e manaʻo ana e hōʻoia i ke kūlike o nā hopena, ua hana ʻia ka 100,000 manawa kuni mua a me 100,000 Markov Chain Monte Carlo (MCMC) iterations ma lalo o ke kumu hoʻohui a me nā alapine allele kūʻokoʻa i waena o nā heluna. Ua hoʻoholo ʻia ka waiwai K ma ka hoʻokō ʻana i ke ʻano AK, i wehewehe ʻia e [60], ma ka papahana pūnaewele STRUCTURE HARVESTER [61]. Ua hāʻawi ʻia kekahi heluna kanaka i kahi puʻupuʻu kikoʻī inā ʻoi aku ka nui o ka helu o ka lālā (q-value) ma mua o 0.7, inā ʻaʻole i manaʻo ʻia he ʻohana hui.
Ua hana ʻia ka loiloi hoʻonohonoho kumu i mea e ʻike ai i nā ʻano o ka pilina genetic ma waena o nā komo i hōʻike ʻia e ka Nei's genetic distance matrix (Supplementary Table S5). Ma muli o nā alapine allele, ua kūkulu ʻia kahi dendrogram o ka mamao genetic me ka hoʻokō ʻana i ke ʻano hui pūʻulu kaumaha ʻole me ka helu helu helu (UPGMA) cluster analysis ma ka polokalamu POPTREEW. [62]. Ua noi ʻia ʻo Bootstrapping e loiloi i ka hilinaʻi i ka hoʻonohonoho hierarchical, e hoʻonohonoho ana i ka 100 resampling o ka hoʻonohonoho ʻikepili. ʻO ka mea hope loa, ʻo ka polokalamu MEGA X [63] ua hoʻohana ʻia ma ke ʻano he polokalamu kiʻi lāʻau.
Nā mea kōkua: Aia ka mea ma ʻaneʻi ma ka pūnaewele http://www.mdpi.com/2223-7747/9/2/260/s1. Pākuʻi S1: Hōʻikeʻike moʻokalaleo o nā puʻupuʻu ARO, MCO, a me TRO. Papa S2: Heterozygosity a me ka ho'opa'a 'ana i helu 'ia no ARO landraces a TRO a me MCO landraces. Pākuʻi S3: Nā waiwai paʻa o ka ʻāpana Fpt. Papa S4: Ka papa inoa o nā SSR i hoʻohana ʻia ma ke aʻo ʻana. Papa S5. Pairwise population matrix of Nei genetic distance. Kiʻi S1: Hoʻololi ka pakuhi laina o nā waiwai K me ka Delta K o Evanno.
Mea kākau Contributions: Ua hāpai ʻo CL a me LR i ke aʻo ʻana a hoʻolālā i ka hoʻokolohua; Ua hana ʻo CL a me PI i ka nānā ʻana i ka māka molekala; Ua hana ʻo ARM a me VZ i nā hoʻokolohua kahua; Ua komo ʻo RM, SP, GR, a me CL i ka ʻikepili ʻikepili; Ua kākau ʻo RM lāua ʻo CL i ka palapala. Ua heluhelu a ʻae nā mea kākau a pau i ka mana i paʻi ʻia o ka manuscript.
Kākoʻo kālā: Ua kākoʻo ʻia kēia hana e ka papahana Regional Apulian "Biodiversity of Apulian vegetable species"—Programma di Sviluppo Rurale per la Puglia 2014-2020. Misura 10—Sottomisura 10.2; hāʻawi i ka CUP H92C15000270002, Italia.
'Ōlelo hōʻeia: Loaʻa nā mahalo iā "Azienda Agricola Iannone Anna" a me "Associazione produttori della vera cipolla rossa di Acquaviva" no ka hoʻolako ʻana i nā mea kanu i hoʻohana ʻia i ka hoʻokolohua.
Nā paio o ka lokomaikaʻi: ʻAʻole i hōʻikeʻia e nā mea kākau i ka paʻi kālā.
E hoʻomaopopo '
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