Luigi Ricciardi 1 , Rosa Mazzeo 2,* © , Angelo Raffaha Marcotrigiano 1 , Guglielmo Rainaldi 3 , Paolo Iovieno 4 , Vito Zono 1 , Stefano Pavan 1© na Concetta Lotti 2,*
- 1 Ngalaba nke ala, ihe ọkụkụ na sayensị nri, mkpụrụ ndụ ihe ọkụkụ na ngalaba ọmụmụ University nke Bari, Via Amendola 165/A, 70125 Bari, Italy; luigi.ricciardi@uniba.it (LR);angelo.marcotrigiano@uniba.it (ARM); vito.zonno@uniba.it (VZ); stefano.pavan@uniba.it (SP)
- 2 Ngalaba Sayensị nke Agriculture, nri na gburugburu ebe obibi, Mahadum Foggia, Via Napoli 25, 71122 Foggia, Italy
- 3 Ngalaba Biosciences, Biotechnologies na Biopharmaceuticals, Mahadum nke Bari, Via Orabona 4, 70125 Bari, Italy; guglielmo.rainaldi@uniba.it
- 4 Ngalaba Teknụzụ Ike, Bioenergy, Biorefinery na Green Chemistry Division, ENEA Trisaia Research Center, SS 106 Ionica, km 419 + 500, 75026 Rotondella (MT), Italy; paolo.iovieno@enea.it
* Mmekọahụ: rosa.mazzeo@unifg.it (RM); concetta.lotti@unifg.it (CL)
Abstract:
Yabasị (Allium cepa L.) bụ ihe ọkụkụ nke abụọ kacha mkpa n'ụwa niile, a na-enwekwa ekele maka uru ahụike ya. N'agbanyeghị mkpa akụ na ụba ya dị mkpa yana uru ọ bara dị ka nri na-arụ ọrụ, yabasị enyochabeghị nke ọma gbasara ụdị mkpụrụ ndụ ihe nketa ya. N'ebe a, anyị nyochara ụdị mkpụrụ ndụ ihe nketa dị na "Acquaviva red yabasị" (ARO), bụ ala nke nwere akụkọ ihe mere eme nke narị afọ gara aga na obere obodo dị na mpaghara Bari (Apulia, Southern Italy). A na-eji ihe nrịbama microsatellite 11 iji chọpụta mgbanwe mkpụrụ ndụ ihe nketa na mkpokọta germplasm nke nwere ọnụ ọgụgụ 13 ARO na ụdị azụmahịa atọ. Nyocha nke usoro mkpụrụ ndụ ihe nketa na usoro parametric na nke na-abụghị parametric mere ka ọ pụta ìhè na ARO na-anọchite anya ọdọ mmiri a kọwara nke ọma, nke dị iche na Tropea na Montoro landraces nke a na-ehiekarị ụzọ. Iji nye nkọwa nke bulbs, nke a na-ejikarị eme ihe maka oriri ọhụrụ, a na-enyocha ọdịnaya siri ike na nke siri ike, na-egosi uto dị elu na ARO n'ihe gbasara ala abụọ a kpọtụrụ aha n'elu. N'ozuzu, ọmụmụ ihe dị ugbu a bara uru maka ọdịnihu valorization nke ARO, nke nwere ike ịkwalite site na akara ngosi dị mma nke nwere ike inye aka belata aghụghọ azụmahịa ma melite ego nke ndị obere.
Okwu Mmalite
Ụdị ụdị Allium gụnyere ihe dị ka ụdị 750 [1], n'ime ya yabasị (Allium cepa L., 2n = 2x = 16) bụ otu n'ime ihe ndị kacha gbasaa. A. cepa nwere okirikiri biennial yana omume ịmụ nwa. N'oge a, mmepụta yabasị zuru ụwa ọnụ (97.9 Mt) na-eme ka ọ bụrụ ihe ọkụkụ nke abụọ kachasị mkpa na-esote tomato [2]. Kemgbe oge ochie, a na-eji bọlbụ yabasị ma dị ka nri ma na ọgwụ ndị mmadụ. N'ezie, ndị Ijipt oge ochie akọworị ọtụtụ usoro ọgwụgwọ dabere na iji galik na yabasị na papyrus ahụike nke 1550 BC, Codex Ebers [3].
A na-eri akwụkwọ nri a na-agbanwe agbanwe ma dị mma nke ọma, nke dị ọhụrụ, ma ọ bụ dị ka ngwaahịa a na-edozi, ma jiri ya mee ka uto nke ọtụtụ efere dịkwuo ụtọ. Ọtụtụ nnyocha e mere n'oge na-adịbeghị anya na-ekwu na ịṅụ yabasị nwere ike ibelata ihe ize ndụ nke ọrịa obi [4,5], oke ibu [6], ọrịa shuga [7], na ụdị ọrịa cancer dị iche iche [8-10]. A na-ekwukarị uru ahụike yabasị na ọkwa dị elu nke klaasị abụọ nke ogige nutraceutical: flavonoids na alk (en) yl cysteine sulphoxides (ACSOs). Klas nke mbụ gụnyere flavonols na anthocyanins. Quercetin bụ isi flavonol a na-achọpụta, mara maka ike antioxidant siri ike na ihe mgbochi mkpali na mkpochapụ radical free na ngbanwe metal ion. [11]; ebe anthocyanins na-enye agba uhie/acha odo odo nye ụfọdụ ụdị yabasị. Banyere ACSO, nke kachasị ukwuu bụ isoalliin [(+) -trans-S-1-propenyl-L-cysteine sulfoxide] [12], sọlfọ amino acid na-abụghị nke na-adịghị agbanwe agbanwe na nke na-abụghị proteinogenic echekwara na sel, bụ nke na-ahụ maka isi ísì ụtọ na uto yabasị n'ụzọ na-apụtaghị ìhè. [13]. Mgbe anụ ahụ mebiri emebi, enzyme alliinase na-agbaji isoalliin ka ọ mepụta ọtụtụ ogige na-agbanwe agbanwe (pyruvate, amonia, thiosulphonates na propanethial S-oxide) nke na-akpali ịdọka ma na-ebute isi na-adịghị mma. [14]. A na-atụkarị ntụpọ yabasị dị ka ọnụọgụ, kwa gram nke ịdị arọ ọhụrụ, nke pyruvic acid nke hydrolysis mepụtara. [15,16].
Na mba nke Mediterranean efere, chọrọ dị ka otu n'ime nke abụọ di iche iche emmepe nke A. cepa [17,18], bọlbụ yabasị na-egosiputa mgbanwe dị ukwuu n'ụdị, nha, agba, ihe akọrọ, na nkwụ [19-22]. Ọzọkwa, fatịlaịza sitere na sọlfọ, omume agronomic, ụdị ala, ọnọdụ ihu igwe, na genotype nke cultivars ma ọ bụ ala nwere ike imetụta àgwà bọlbụ site n'inye ụkpụrụ organoleptic pụrụ iche na nri nri. [23-27]. Na Italy, n'agbanyeghị nnukwu yabasị germplasm dị, ọ bụ naanị ụdị yabasị ole na ole ka a na-edobe ọmụmụ sayensị ma mara nke ọma. [28,29].
Nkọwapụta nke ọma mkpụrụ ndụ ihe nketa na phenotypic nke agro-bidiversity dị oke mkpa iji hụ na nchekwa nchekwa mkpụrụ ndụ ihe ọkụkụ kwesịrị ekwesị ma kwalite ojiji nke ụdị mkpụrụ ndụ ihe nketa akọwapụtara n'agbụ ọnụ ahịa. [30-32]. A na-ahọrọkarị nrịbama usoro nkwugharị dị mfe (SSR) maka eserese [33-35], Mkpịsị aka DNA na ịkpa oke cultivar [36-38], na ntule ntụkwasị obi nke mgbanwe mkpụrụ ndụ ihe nketa n'ime na n'etiti agbụrụ ala [39-42], ebe ọ bụ na ha bụ kpọmkwem ebe, multi-allelic, codeominatly ketara, ukwuu reproducible, na dabara maka akpaghị aka genotyping.
N'ime ọmụmụ ihe ugbu a, anyị lekwasịrị anya n'okirikiri ọdịnala Apulian, "Acquaviva red onion" (ARO), nke a na-akọ dị ka usoro ọrụ ugbo si dị na obere mpaghara nke obodo Acquaviva delle Fonti, na mpaghara Bari. (Apulia, Southern Italy). Okooko osisi nke ala ala a buru ibu ma gbawara agbawa na agba ọbara ọbara ma bụrụ nke a na-ejikarị eme ihe na ntụziaka mpaghara. Agbanyeghi na ARO nwetara akara ngosi “Slow Food Presidium”, enwere ike ịkwalite ma chekwaa mmepụta ya site na akara mma European Union dị ka ihe nrịbama ala echedoro (PGI) na aha echekwabara (POD), n'ihi na ndị a nwere ike inye aka belata aghụghọ azụmahịa na imeziwanye ego nke ndị nwe obere. N'ime ebe a, ejiri akara molecular SSR mee ihe dị ka ngwa ọrụ siri ike iji chọpụta mgbanwe mkpụrụ ndụ ihe nketa n'etiti ndị ARO yana ịkpa oke ala a site na agbụrụ abụọ na-acha uhie uhie nke Southern Italian. Ọzọkwa, anyị mere atụmatụ pungency na soluble ọdịnaya siri ike iji nyochaa ụtọ ARO n'ihe metụtara ahịa ahịa.
Results
Nhazi nke Acquaviva Red yabasị Germplasm mkpokọta na njirimara ọdịdị ọdịdị
Mkpụrụ nke mmadụ iri na atọ nke ARO landrace, nke ndị ọrụ ugbo nyere n'usoro ọrụ BiodiverSO Apulia Region were guzobe mkpokọta germplasm ARO.
Achịkọtara nkọwa gbasara ọdịdị ọdịdị ọdịdị, nke metụtara bọlbụ, akpụkpọ anụ, na anụ ahụ na ARO germplasm na n'elu ala yabasị atọ, abụọ bụ nke "Tropea red yabasị" (TRO) landrace na otu nke "Montoro ọla kọpa yabasị" (MCO) landrace (Ọgụgụ). 1). Igwe ọkụ ARO niile dị larịị ma bụrụ nke e ji mara akpụkpọ ahụ na-acha uhie uhie na mpụga nke nwere ndo uhie dị iche iche. N'ụzọ dị iche, anụ ahụ nke TRO bulbs na-acha uhie uhie zuru oke, ebe anụ ahụ nke MCO bulbs na-adịghị mma pigmented (Table S1). Nyocha biochemical kwere ka nyochaa ọdịnaya siri ike soluble yana nhụsianya. Dị ka a kọrọ na Tebụl 1, Ntụle ụkpụrụ nke ọdịnaya siri ike soluble nke bulbs na ndị ARO bụ 7.60, wee malite site na 6.00 (ARO12) ruo 9.50° Brix (ARO11 na ARO13). Uru a dị elu karịa nke e mere atụmatụ maka ala ndị TRO na MCO (4.25 na 6.00° Brix, n'otu n'otu).
Ntọala 1. Ọdịnaya Soluble siri ike na uru bara uru A tụlere na "Acquaviva Red yabasị" (ARO), "Tropea Red yabasị" (TRO), na "Montoro Copper Onion" (MCO) *.
CODE | Ọdịnaya siri ike soluble (Brix) | Obere oge (pmolg-1 FW) | ||
Ọ pụtara | CV y (%) | Ọ pụtara | CV y (%) | |
ARO1 | 6.25 D * | 5.65 | 5.84 ab * | 23.78 |
ARO2 | 7.25 DC | 4.87 | 6.51 ka | 22.98 |
ARO3 | 7.50 BCD | 9.42 | 5.28 ab | 22.88 |
ARO4 | 7.50 BCD | 0.00 | 6.97 ka | 3.74 |
ARO 5 | 7.50 BCD | 0.00 | 6.80 ka | 9.68 |
ARO6 | 6.25 D | 5.65 | 4.51 ab | 39.18 |
ARO7 | 7.25 DC | 4.87 | 5.25 ab | 15.44 |
ARO8 | 9.00 AB | 0.00 | 7.04 ka | 3.49 |
ARO9 | 8.25 ABC | 4.28 | 6.84 ka | 0.15 |
ARO10 | 7.00 DC | 0.00 | 5.94 ab | 6.57 |
ARO11 | 9.50 A | 7.44 | 5.54 ab | 16.43 |
ARO12 | 6.00 D | 0.00 | 4.91 ab | 9.70 |
ARO13 | 9.50 A | 7.44 | 6.63 ka | 24.93 |
MCO | 6.00 D | 0.00 | 4.18 ab | 2.66 |
TRO1 | 4.25 E | 8.31 | 2.80 nk | 2.10 |
TRO2 | 4.25 E | 8.31 | 4.28 ab | 4.79 |
* Pụtara na otu mkpụrụedemede na mkpụrụedemede ukwu ma ọ bụ obere abụghị dị iche iche na 0.01P ma ọ bụ 0.05P, n'otu n'otu (Nnwale SNK). y Ọnụọgụ nke mgbanwe.
Ọnụ ego nke ARO pungency, nke a na-enyocha site na ọdịnaya pyruvic acid, bụ 6.00, sitere na 4.51 pmol g.-1 FW (ARO6) ruo 7.04 (ARO8). Ọnụ ahịa a dị elu karịa nke e mere atụmatụ na TRO na MCO landraces (3.54 pmol g-1 FW na 4.18 pmol g-1 FW, n'otu n'otu).
SSR Polymorphism na Mmekọrịta Genetic n'etiti Accessions
N'ime ọmụmụ ihe ugbu a, 11 n'ime 37 nwalere SSR primer ngwakọta nyere otu-locus polymorphisms, ya bụ, na-ekweta na kacha abụọ ngwaahịa amplification n'ime otu onye. N'ozuzu, achọpụtara 55 alleles n'ime mmadụ 320 nwere ọnụ ọgụgụ alleles kwa mpaghara sitere na 2 (ACM147 na ACM 504) ruo 11 (ACM132) yana uru nke 5 alleles (Table). 2). N'ime ndị mmadụ n'otu n'otu, ọnụ ọgụgụ alleles (Na) sitere na 1.94 (ACM147 na ACM504) ruo 5.38 (ACM132), ebe ọnụọgụ alleles (Ne) dị irè sitere na 1.41 (ACM152) ruo 2.82 (ACM449). Ndịiche n'etiti ụkpụrụ Na na Ne bụ n'ihi ọnụnọ alleles nwere obere oge na ọnụ ọgụgụ ndị mmadụ na oke nke naanị alleles ole na ole. E gosipụtara uru heterozygosity (Ho) kachasị elu maka ACM138 na ACM449 (0.62), ebe nke kacha ala jikọtara ya na ACM152 (0.25). Atụrụ anya heterozygosity (He), nke kwekọrọ na atụmanya echiche na ọnụ ọgụgụ panmictic, sitere na 0.37 (ACM504) ruo 0.61 (ACM132, ACM138, na ACM449). Wright's fixation index (Fis), gosipụtara ụkpụrụ dị nso na efu (nkezi 0.05) maka akara niile, na-egosi ụkpụrụ ndị yiri ya n'etiti ọkwa heterozygosity hụrụ na a tụrụ anya ya, dịka a tụrụ anya maka ụdị na-agafe. A na-eme atụmatụ arụmọrụ nke ihe nrịbama SSR n'otu n'otu na mkpịsị aka mkpụrụ ndụ ihe nketa site na ndeksi ọdịnaya ozi polymorphic (PIC), yana uru pụtara 0.48 wee malite na 0.33 (ACM504) ruo 0.67 (ACM132). Ndekọ arụmọrụ ọzọ, Index Ozi Shannon (I) gosipụtara uru pụtara 0.84, wee were ụkpụrụ sitere na 0.45 (ACM152) ruo 1.20 (ACM132).
Ntọala 2. Akụkụ Polymorphism nke 11 SSR nrịbama ejiri iji tụọ ụdị dị iche iche nke mkpụrụ ndụ ihe nketa na ARO, TRO, na MCO. Ngụkọta ọnụ ọgụgụ Alleles (Na), Oke Band Size, na Polymorphic Ozi Ọdịnaya (PIC) ntụaka na-ezo aka na mkpokọta ndị mmadụ 320 Genotyped na ọmụmụ ihe a. Ọnụọgụ Alleles (Na), ọnụ ọgụgụ Alleles dị irè (Ne), Heterozygosity hụrụ (Ho), Heterozygosity tụrụ anya (He), Index Fixation (F)is), na Shannon's Information Index (I) na-ezo aka n'ụkpụrụ ndị a gbakọrọ site na ọnụ ọgụgụ mmadụ 16, nke ndị mmadụ 20 mejupụtara nke ọ bụla.
Mpaghara. | Mkpokọta Na | Oke oke (bp) | eserese | Ọ pụtara | |||||
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 |
Ọ pụtara | 5 | 0.48 | 3.22 | 2.15 | 0.46 | 0.48 | 0.84 | 0.05 |
N'ime ọnụ ọgụgụ ndị mmadụ, ARO3, ARO6, ARO8, ARO10, TRO1, na MCO gosipụtara mgbanwe dị elu nke mkpụrụ ndụ ihe nketa (Ho> 0.5), ebe a na-ahụ ụdị dị iche iche nke dị ala na ndị bi na ARO7 (Ho = 0.27) (Spplementary Table S2). N'ozuzu, ntinye niile gosipụtara Fis ụkpụrụ dị nso na efu (Fis uru pụtara = 0.054), dị ka a tụrụ anya n'okpuru ọnọdụ mating random.
Nyocha nke ọdịiche Molecular na Ọdịdị mkpụrụ ndụ ihe nketa
AMOVA gbakọtara nkewa nhazi usoro mgbanwe mkpụrụ ndụ ihe nketa n'etiti na n'ime ndị mmadụ. Nsonaazụ a gosiputara obere akụkụ nke mgbanwe mkpụrụ ndụ ihe nketa n'ime ndị mmadụ (pasent 87). Ọdịiche dị n'etiti ndị mmadụ, 13%, dị oke mkpa (P <0.001) (Obodo 3). Ụkpụrụ ụzọ abụọ nke paramita Fpt, ihe yiri nke Wright's Fst fixation index, sitere na 0.002 (ARO2/ARO10) ruo 0.468 (ARO7/TRO2), dị ịrịba ama (P <0.05), ewezuga ntụnyere ụzọ itoolu (Table S3).
Ntọala 3. Nyocha nke Molecular Variance nke 320 Genotypes si 16 Ọnụ ọgụgụ nke Allium akwa L.
Source | df | Mgbakọ nke square | Ntụle dị iche iche | Ndịiche (%) | Fpt | P |
N'etiti ndị mmadụ | 15 | 458.63 | 1.16 | 13% | ||
N'ime ọnụ ọgụgụ mmadụ | 304 | 2272.99 | 7.50 | 87% | 0.134 | 0.001 |
Total | 319 | 2731.62 | 8.66 |
Nnyocha nke usoro mkpụrụ ndụ ihe nketa na A. cepa A na-eme mkpokọta genotyped n'ime ọmụmụ ihe a site na nlebanya nchịkọta nke gbanyere mkpọrọgwụ nke etinyere n'ọrụ n'ụdị ngwanro. Ụzọ Evanno AK tụrụ aro nkewa n'ụyọkọ abụọ (K = 2) dị ka ihe kacha enye nkọwa maka anyị. ihe omuma,ya na nkee Osote kacha peaka na K = 5 (nsopplementaiv Rgure OBI). A maka K = 2, ahpopiwu were agbaigned na onnaf ụyọkọ abụọ ahụ na ọnụọgụ rnernbertoip (q)> 0.7. Dị ka shown na ọnụ ọgụgụ 2a, ụyọkọ nke mbụ (aha ya bụ S1) gụnyere MCO na ndị ARO niile, ebe ụyọkọ S2 chịkọtara ọnụ ọgụgụ TRO abụọ ahụ. Na K = 5, na-enye nkọwa miri emi nke dataset (Fig 2b), 75% nke ntinye ka ekenyere otu n'ime ụyọkọ ise ahụ. A kwadoro nkewa n'etiti ARO (S1) na TRO (S2), n'agbanyeghị na ejikọtara ụfọdụ ndị ARO (q <0.7) ma ọ bụ jikọta iche na ụyọkọ ọhụrụ abụọ S3 na S4 (ARO7 na ARO12, otu). N'ụzọ na-akpali mmasị, ụdị azụmahịa MCO guzobere ụyọkọ dị iche iche (S5) nke kewapụrụ na yabasị uhie Apulian.
Mmekọrịta mkpụrụ ndụ ihe nketa n'etiti ndị mmadụ
SSR polymorphism kwere ka ise dendrogram nke mkpụrụ ndụ ihe nketa di iche iche na nsonaazụ nke phylogenetic analysis na-egosi na Figure. 3a. N'ebe a, e kewara mkpokọta germplasm n'ìgwè ise nke ụkpụrụ bootstrap kwadoro nke ọma. Ndị ARO7 na ARO12 kewapụrụ ozugbo na ndị fọdụrụ wee mepụta ụyọkọ abụọ dị iche iche. Ụyọkọ nke atọ gụnyere ọnụọgụ ahịa abụọ nke TRO, ebe ọnụ ụzọ anọ kewara MCO site na ndị ARO iri na otu. Achọpụtara mmekọrịta mkpụrụ ndụ ihe nketa na-eme n'etiti ndị mmadụ site na nyocha nhazi nhazi isi (PCoA) (Ọgụgụ). 3b). Dịka e gosipụtara na mbụ, a chịkọtara ọnụ ọgụgụ ndị ARO nke ọma, ma e wezụga ARO12 na ARO7, ndị pụtara n'ọnọdụ dịpụrụ adịpụ na atụmatụ PCoA. Ndị TRO abụọ na ndị MCO gbasasịa n'akụkụ aka nri ala nke atụmatụ ahụ.
Nyocha 3. Ụdị mkpụrụ ndụ ihe nketa dị iche iche n'etiti mmadụ iri na isii A. cepa ọnụ ọgụgụ ndị e ji mara na ọmụmụ ihe a, dabere na profaịlụ SSR ha. (a) UPGMA dendrogram nke anya mkpụrụ ndụ ihe nketa. A na-egosi ụkpụrụ nkwado bootstrap> 50 n'elu ọnụ ọnụ kwekọrọ; (b) Ntụle akụrụngwa nke isi (PCoA). Ụyọkọ a gbara okirikiri na-acha uhie uhie dabara n'ụzọ zuru oke yana otu ndị nyocha phylogenetic mebere yana nke 11 ARO mebere.
Nkwurịta
N'ime ọnụ ọgụgụ dị ukwuu nke agro-bidiversity nke omenala na-akọ na Southern Italy, yabasị landraces na-anọchi anya ngwaahịa niche dị mkpa ka e chebe ya pụọ n'ihe ize ndụ nke mkpụrụ ndụ ihe mgbochi mbuze na iyi egwu nke nnọchi site n'oge a cultivars. N'ime usoro ọrụ mpaghara BiodiverSO, nke ezubere iji nakọta, ịkọwapụta, ịkwalite na ichekwa akụrụngwa mkpụrụ ndụ ihe nketa nke mpaghara Apulia nwere njikọ siri ike na ihe nketa mpaghara, anyị hibere mkpokọta mkpụrụ nke mmadụ iri na atọ nke ARO landrace. Anyị kọrọ nyocha mbụ nke mgbanwe ARO n'ihe gbasara DNA polymorphisms na abụọ biochemical parameters, soluble solid na pyruvic acid ọdịnaya, metụtara ekpomeekpo àgwà na mkpa maka ịnakwere nke ọhụrụ na-adịghị esi nri ngwaahịa. Na mgbakwunye, e jiri data dị na ala ARO tụnyere nke anakọtara na mba abụọ ọzọ nwere pigmented landraces nke ọ na-emehiekarị.
Nyocha biochemical mere ka ọ pụta ìhè ụtọ nke ndị ARO 13, metụtara ọdịnaya siri ike soluble dị elu na obere oge, dịka ntuziaka ụlọ ọrụ yabasị dị ụtọ si dị. [31]. Bọlbụ ARO dị ụtọ karịa nke ndị TRO na MCO landraces, ma gosipụta obere nhịahụ dị elu. Otú ọ dị, ụtọ na eyịm bụ n'ihi nguzozi n'etiti ọdịnaya shuga na pungency, ya mere njirimara a nwere ike ịba uru iji kwado nhọrọ nke genotypes nke uru, nke ndị ọrụ ugbo na-emekarị na-adabere na morphology.
Ekwenyere akara SSR dị ka ngwá ọrụ bara uru iji kpachapụ anya genotypes, n'agbanyeghị na anakọtara n'ime mpaghara dị warara na-eto eto dị ka obodo Acquaviva delle Fonti. Ihe nrịbama ahọpụtara gosipụtara ọnụ ọgụgụ alleles dị elu karịa akara akara ndị akọpụtara na mbụ [43] na [44], mana ọ dị ala karịa akara ndị a kọrọ [45]. Ọzọkwa, 50% nke ihe nrịbama anyị gosipụtara ụkpụrụ index PIC karịrị 0.5, na-egosi na ọ dabara adaba ịkpa ókè agbụrụ ndị nọ na mkpokọta ahụ, dị ka atụpụtara ya. [46]. Ntụle dị iche iche n'ime ọnụ ọgụgụ mmadụ gosipụtara ụkpụrụ ndị yiri ya n'etiti Ho na He, na-ebute Fi oberes ụkpụrụ. Nke a bụ nkwekọrịta na si-agafe ọdịdị nke A. na, nke na-enwe nnukwu ịda mbà n'obi [47]. N'ozuzu Fis ọnụ ahịa a gbakọrọ na ọnụọgụ yabasị tụlere n'ọmụmụ ihe a (0.054) dị ala karịa nke nke a kọrọ na mbụ. [45] (0.22) na ihe fọrọ nke nta ka ọ bụrụ nke ahụ hụrụ [31] (0.08) na [48] (0.00) onye nyochara ụdị mkpụrụ ndụ ihe nketa dị iche iche na ala yabasị sitere na ugwu ọdịda anyanwụ Spain na Niger, n'otu n'otu. Ọkwa dị iche iche nke heterozygosity dị na ndị ARO na-eme ka echiche ahụ na Apulia na-anọchite anya ebe dị iche iche maka ọtụtụ ụdị osisi. [32, 42, 49-51].
AMOVA mere ka ọ pụta ìhè na ọtụtụ mgbanwe molekụla na mkpokọta genotyped na ọmụmụ a dị n'ime ndị mmadụ. Agbanyeghị, nnukwu ọdịiche mkpụrụ ndụ ihe nketa n'etiti ndị mmadụ (FPT ụkpụrụ) kpughere na omume nke mkpụrụ ndụ ihe nketa stratification. N'ezie, ọ bụ ezie na nsonaazụ anyị gosipụtara ọnụnọ nke otu mkpụrụ ndụ ihe nketa n'ọtụtụ ndị ARO, na-etolite ụyọkọ akọwapụtara nke ọma, ndị ARO7 na ARO12 gosipụtara profaịlụ mkpụrụ ndụ ihe nketa doro anya. Nsonaazụ a nwere ike ịbụ n'ihi ụzọ dị iche iche sitere na mkpụrụ nke ndị ọrụ ugbo abụọ ahụ na-esi na ya nweta ndị mmadụ. Ọzọkwa, dabere na nsonaazụ enwetara, ARO landrace nwere ike ịtụle nke ọma na ọkwa mkpụrụ ndụ ihe nketa site na ala TRO na MCO. N'ime nnyocha e mere na nso nso a, [29] enyochala ụdị mkpụrụ ndụ ihe nketa dị iche iche nke mba Italy dị iche iche gụnyere "Acquaviva," "Tropea," na "Montoro." Ọ bụ ezie na ndị na-ede akwụkwọ na-eji akara SNP iji chọpụta ụdị mkpụrụ ndụ ihe nketa dị iche iche nke nchịkọta yabasị sara mbara, genotyping enweghị ike ịkpa ókè "Acquaviva" sitere na eyịm "Tropea" na "Montoro". Eleghị anya, nghọtahie a bụ n'ihi ọnụ ahịa PIC dị ala achọtara (0.292), na-atụ aro obere ozi izugbe nke ebe a na-enyocha dị ka a na-ekwu. [29]. Ọzọkwa, iji nyochaa ọnụnọ nke sub-structure na ụyọkọ Ịtali ha, ọ ga-aka mma iji nyochaa genotypes Ịtali dị iche iche na nchịkọta ndị ọzọ. Eleghị anya ọ gaara ekwe ka iji anya nke uche hụ ụkpụrụ nke ụdị mkpụrụ ndụ ihe nketa dị iche iche jikọtara ya na nhazi ala ma ọ bụ àgwà dị n'okpuru nhọpụta siri ike.
N'ikpeazụ, ọmụmụ ihe dị ugbu a na-anọchite anya akụkọ zuru oke gbasara agbụrụ ala yabasị jikọtara ya na omenala omenala obodo na mkpa akụ na ụba maka ndị ọrụ ugbo. Nsonaazụ anyị na-eme ka ọ pụta ìhè na, ma e wezụga ihe ole na ole, ARO na-eji ọdọ mmiri a na-akọwa nke ọma, bụ nke kwesịrị ka e chebe ya pụọ na ihe ize ndụ nke mmebi mkpụrụ ndụ ihe nketa. Ya mere, nguzobe nke nchịkọta nnọchiteanya nke isi iyi dị iche iche nke mkpụrụ ndụ ihe nketa dị oké mkpa. N'ikpeazụ, njirimara mkpụrụ ndụ ihe nketa na phenotypic nke ARO nwere ike ịba uru iji nweta akara ngosi n'aka European Union.
Akụrụngwa na ụzọ
Mkpokọta Germplasm, ihe ọkụkụ, na mmịpụta DNA
E nwetara ọtụtụ mmadụ iri na atọ nke ARO landrace n'ime usoro nke ọrụ mpaghara Apulia (BiodiverSO: https://www.biodiversitapuglia.it/), site n'usoro ozi nke emere na "Acquaviva delle Fonti", obere obodo Apulian dị na mpaghara Bari, Italy. Edebere saịtị mkpokọta nke ohere ọ bụla site na Sistemụ Ozi Geographic (GIS) wee kọwapụta ya na Tebụl 4. Na mgbakwunye, ọnụ ọgụgụ mmadụ abụọ sitere na TRO landrace na otu ndị sitere na MCO landrace gụnyere n'ọmụmụ ihe ugbu a ma jiri ya mee ihe dị ka ntụaka. A na-akụ ihe ọkụkụ niile n'otu ọnọdụ gburugburu ebe obibi na ugbo nnwale "P Martucci" nke Mahadum Bari (41° 1'22.08″ N, 16°54'25.95″ E), n'okpuru mkpuchi iji zere ịgbasa pollination n'etiti. ọnụ ọgụgụ ndị mmadụ na-emesi ike na mmetọ n'ime mmadụ site na ijiji (Lucilia Caesar). E ji mara ndị mmadụ iri na isii ahụ maka àgwà ndị metụtara nha bọlbụ na udi na agba anụ ahụ (Table S16). Na mgbakwunye, a na-eme nyocha ọdịnaya siri ike soluble site na iji refractometer ejiri aka wee tụọ nhụsianya na nlele ihe ọṅụṅụ yabasị na-agbakwunye 1-dinitrophenyl hydrazine (2,4% v/v na 2N nke HCl) na nyochaa nnabata na 420 nm, dị ka akọwara [31]. Emere ule ọtụtụ oke Duncan na ule SNK iji chọpụta ọnụnọ dị iche iche.
Ntọala 4. Ndepụta ọnụ ọgụgụ ndị anakọtara na Genotyped na ọmụmụ ihe a. Maka ọnụ ọgụgụ mmadụ ọ bụla, koodu njirimara, aha mpaghara, GPS Coordinate, na Gene Bank na-echekwa mkpụrụ osisi ka akọpụtala.
Usoro | aha | Ndị na -ahazi GPS | Ụlọ ọrụ Gene Bank y |
ARO1 | Cipolla rossa di Acquaviva | 40°54’21.708″ N 16°49’1.631” E | Di.SSPA |
ARO2 | Cipolla rossa di Acquaviva | 40°53’14.28″ N 16°48’56.879” E | Di.SSPA |
ARO3 | Cipolla rossa di Acquaviva | 40°54’11.304″ N 16°49’13.079” E | Di.SSPA |
ARO4 | Cipolla rossa di Acquaviva | 40°54’3.348″ N 16°40’27.011” E | Di.SSPA |
ARO5 | Cipolla rossa di Acquaviva | 40°51’59.76″ N 16°53’0.527” E | Di.SSPA |
ARO6 | Cipolla rossa di Acquaviva | 40°52’48.72″ N 16°49’43.247” E | Di.SSPA |
ARO7 | Cipolla rossa di Acquaviva | 40°53’13.47″ N 16°50’23.783” E | Di.SSPA |
ARO8 | Cipolla rossa di Acquaviva | 40°53’18.816″ N 16°49’33.888” E | Di.SSPA |
ARO9 | Cipolla rossa di Acquaviva | 40°54'51.372 ″ N 16°49'3.504" E | Di.SSPA |
ARO10 | Cipolla rossa di Acquaviva | 40°54’1.188″ N 16°49’24.311” E | Di.SSPA |
ARO11 | Cipolla rossa di Acquaviva | 40°52'49.8 ″ N 16°49'48.575" E | Di.SSPA |
ARO12 | Cipolla rossa di Acquaviva | 40°52’38.892″ N 16°49’28.379” E | Di.SSPA |
ARO13 | Cipolla rossa di Acquaviva | 40°53’21.768″ N 16°49’29.711” E | Di.SSPA |
TRO1 | Cipolla rossa lunga di Tropea | - | Di.SSPA |
TRO2 | Cipolla rossa tonda di Tropea | - | Di.SSPA |
MCO | Cipolla Ramata di Montoro | - | Di.SSPA |
y Di.SSPA, ngalaba nke ala, osisi na sayensị nri, Mahadum Bari. |
A na-enyocha akwụkwọ mpempe akwụkwọ nke 20 genotypes kwa mmadụ ma chekwaa ya na -80 Celsius ruo mgbe ejiri ya. Maka ụdị polysaccharide bara ụba, dịka A. na, Nzọụkwụ mbụ iwepụ polysaccharide dị mkpa iji nweta DNA dị mma, yabụ na-asachapụ mbụ na STE buffer (0.25 M sucrose, 0.03 M Tris, 0.05 M EDTA) dị ka akọwara ya. [52]. Mkpokọta DNA ka ewepụtara na-eso usoro CTAB [53] na n'ikpeazụ, a na-enyocha ya maka ịdị mma na ntinye uche site na Nano Drop 2000 UV-vis spectrophotometer (ThermoScientific, Waltham, MA, USA) na 0.8% agarose gel electrophoresis.
Nyocha SSR
Ngwakọta primer 16 EST-SSR mepụtara site na [54] na mbụ nwalere na mkpụrụ ndụ ihe nketa di iche iche ọmụmụ site [43] na [44] na 21 genomic SSR [45-55] e nyochara ya iji nyochaa ntozu ha (Table S4). Emere Genotyping site na iji usoro mkpado fluorescent akụ na ụba nke etinyere ọdụ M13 na primer SSR ọ bụla na-aga n'ihu. [56]. A kwadebere ngwakọta PCR na mmeghachi omume 20 gL nwere: 50 ng nke DNA zuru ezu, 0.2 mM nke dNTP mix, 1X nke PCR mmeghachi omume buffer, 0.8 U nke DreamTaq DNA polymerase (Thermo Scientific, Waltham, MA, USA), 0.16 gM nke reverse primer. , 0.032 gM nke primer na-aga n'ihu gbatịpụrụ ya na usoro M13 (5'-TGTAAAACGACGGCCAGT-3′), na 0.08 gM nke ihe mkpuchi M13 zuru ụwa ọnụ nke akpọrọ FAM ma ọ bụ NED fluorescent dyes (Sigma-Aldrich, St. Louis, MO, USA). Emere mmeghachi omume PCR na SimpliAmp (Applied Biosystems, CA, USA) thermocycler nwere ọnọdụ ndị a maka ọtụtụ ụzọ abụọ mbụ: 94 Celsius C maka 5 min, okirikiri 40 na 94 Celsius C maka 30 s, 58 Celsius C. maka 45 s na 72 Celsius C maka 45 s na elongation ikpeazụ na 72 Celsius C maka 5 min. Maka ACM446 na ACM449, etinyere PCR mmetụ aka site na mwepu nke 60 Celsius C ruo 55 Celsius C n'elu okirikiri iri, okirikiri 10 na 30 Celsius C, na mgbakwunye ikpeazụ nke 55 min na 5 Celsius C. A na-etinye ngwaahịa PCR n'ime efere 72 nke ọma ma jikọta ya na 96 gL nke Hi-Di Formamide (Life Technologies, Carlsbad, CA, USA) na 14 gL GeneScan 0.5 ROX Size Standard (Life Technologies, Carlsbad, CA, USA). Emeziri amplicons site na ABI PRISM 500 Avant Genetic Analyzer (Life Technologies, Carlsbad, CA, USA) igwe sequencing capillary, ebe akara alleles dị ka onye na-achịkọta ma kenye ya site na iji GeneMapper Software Version 3100.
Ngwa ngwanrọ GenAlEx 6.5 [57] na Cervus 3.0.7 [58] A na-eji ọnụ ọgụgụ nke alleles (Na), ọnụ ọgụgụ nke alleles na-arụ ọrụ (Ne), hụrụ heterozygosity (Ho), heterozygosity na-atụ anya (He), ọdịnaya ozi polymorphic (PIC), nchịkọta ozi Shannon (I), na ntinye ntinye (Fis). ) maka mpaghara SSR ọ bụla.
Ntụle nke ụdịdị mkpụrụ ndụ ihe nketa
GenAlEx 6.5 tụlere ikewa usoro nke usoro mkpụrụ ndụ ihe nketa n'etiti na n'ime ọnụ ọgụgụ yabasị. [57] site na nyocha nke molecular variance (AMOVA) na 999 bootstrapping iji nwalee maka mkpa. Ọzọkwa, a na-eji ngwanrọ GenAlEx 6.5 mee atụmatụ dị iche iche dị n'ime mmadụ ọ bụla site na ịgbakọ nkezi nke Ho, He, na Fis n'elu ebe SSR niile.
Atụmatụ mkpokọta ọnụ ọgụgụ ndị bi na Bayesian bụ ụyọkọ algọridim nke etinyere n'ọrụ na ngwa ngwa STRUCTURE v.2.3.4. [59]. Ejiri ọtụtụ ụyọkọ hypothetical (K), malite na 1 ruo 10, na-esetịpụ ọsọ nke nnwere onwe iri maka uru K nke ọ bụla. Maka ọsọ nke ọ bụla, na-achọ ịchọpụta nkwụsi ike nke nsonaazụ ya, 100,000 mbido ọkụ na oge 100,000 Markov Chain Monte Carlo (MCMC) ka emere n'okpuru usoro nnabata yana ugboro ole na-anọpụ iche n'etiti ndị mmadụ. O yikarịrị ka uru K kpebisiri ike itinye usoro AK, nke kọwara ya [60], n'ime ihe omume webụsaịtị STRUCTURE HARVESTER [61]. E kenyere ọnụ ọgụgụ mmadụ n'otu n'otu n'otu ụyọkọ mgbe ọnụọgụ otu ya (q-uru) dị elu karịa 0.7, ma ọ bụghị ya, a na-ele ya anya dị ka agbụrụ agbakwunyere.
Emere nyocha nhazi nke isi ka iji hụ anya n'ụkpụrụ nke mmekọrịta mkpụrụ ndụ ihe nketa n'etiti njikọ nke Nei's genetic distance matrix (Mgbakwunye Tebụl S5) kpughere. Dabere na frequency allele, a rụrụ dendrogram nke mkpụrụ ndụ ihe nketa na-emejuputa usoro ụzọ otu ụzọ na-enweghị oke yana nyocha ụyọkọ mgbakọ na mwepụ (UPGMA) na ngwanrọ POPTREEW. [62]. A na-etinye ihe nrịbama iji chọpụta ntụkwasị obi dị na nchịkọta ndị isi, na-edozi 100 resampling nke data setịpụrụ. N'ikpeazụ, MEGA X software [63] eji dị ka osisi osise software.
Ngwa mgbakwunye: Ndị na-esonụ dị online na http://www.mdpi.com/2223-7747/9/2/260/s1. Tebụl S1: Nkọwapụta gbasara mmụọ nke ARO, MCO, na TRO bulbs. Tebụl S2: Heterozygosity na ndenye ndozi gbakọọ maka ala ala ARO na ala TRO na MCO. Tebụl S3: Ụkpụrụ ụzọ abụọ nke oke Fpt. Tebụl S4: Ndepụta nke SSR ejiri mee ihe n'ọmụmụ ihe. Tebụl S5. Matriks ọnụọgụ ọnụọgụ abụọ nke ogologo mkpụrụ ndụ ihe nketa Nei. Ọgụgụ S1: eserese ahịrị nke ụkpụrụ K na-agbanwe na Evanno's Delta K.
Onye nyere onyinye: CL na LR chepụtara ọmụmụ ihe ahụ wee chepụta nnwale ahụ; CL na PI mere nyocha ihe nrịbama mkpụrụ ndụ; ARM na VZ mere ule ubi; RM, SP, GR, na CL tinyere aka na nyocha data; RM na CL dere ihe odide a. Ndị ode akwụkwọ niile agụọla ma kwenye na ụdị ihe odide ahụ ebipụtara.
Ego: Ego a na-arụ ọrụ a bụ Regional Apulian Project "Bidiversity of Apulian akwukwo nri ụdị" -Programma di Sviluppo Rurale per la Puglia 2014-2020. Misura 10—Sottomisura 10.2; nye CUP H92C15000270002, Italy.
Nkweta: Ekele bụ n'ihi "Azienda Agricola Iannone Anna" na "Associazione produttori della vera cipolla rossa di Acquaviva" maka ịnye ihe ọkụkụ eji eme ihe na nnwale ahụ.
Esemokwu Mmasị: Ndị na-ede akwụkwọ ekwupụtaghị mmasị ọ bụla.
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