Luigi Ricciardi 1 , Rosa Mazzeo 2,*© , Angelo RaffAele Marcotrigiano 1 , Guglielmo Rainaldi 3 , Paolo Iovieno 4 , Vito Zonno 1 , Stefano Pavan 1© le Concetta Lotti 2,*
- 1 Lefapha la Mobu, Lisaense tsa Limela le Lijo, Setsi sa Liphatsa tsa lefutso tsa limela le ho tsoala Univesithi ea 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 Lefapha la Saense ea Temo, Lijo le Tikoloho, Univesithi ea Foggia, Via Napoli 25, 71122 Foggia, Italy
- 3 Lefapha la Bioscience, Biotechnologies le Biopharmaceuticals, Univesithi ea Bari, Via Orabona 4, 70125 Bari, Italy; guglielmo.rainaldi@uniba.it
- 4 Lefapha la Matla a Theknoloji, Bioenergy, Biorefinery le Green Chemistry Division, ENEA Trisaia Research Center, SS 106 Ionica, km 419 + 500, 75026 Rotondella (MT), Italy; paolo.iovieno@enea.it
* Lengolo: rosa.mazzeo@unifg.it (RM); concetta.lotti@unifg.it (CL)
Sekhetho:
Anyanese (Allium cepa L.) ke sejalo sa bobeli sa bohlokoa sa meroho lefatšeng ka bophara 'me se ananeloa haholo ka melemo ea sona ea bophelo bo botle. Ho sa tsotellehe bohlokoa ba eona ba moruo le boleng ba eona e le lijo tse sebetsang, onion ha e-s'o batlisisoe hantle mabapi le mefuta-futa ea liphatsa tsa lefutso. Mona, re ile ra hlahloba phapang ea liphatsa tsa lefutso ho "Acquaviva red onion" (ARO), sebaka sa naha se nang le histori ea lilemo tse lekholo tsa temo motseng o monyenyane profinseng ea Bari (Apulia, Boroa ho Italy). Ho ile ha sebelisoa sehlopha sa li-microsatellite marker tse 11 ho hlahloba phapang ea liphatsa tsa lefutso ka har'a pokello ea likokoana-hloko e nang le baahi ba 13 ba ARO le mefuta e meraro e tloaelehileng ea khoebo. Litlhahlobo tsa sebopeho sa liphatsa tsa lefutso ka mekhoa ea parametric le e seng ea parametric e totobalitse hore ARO e emela letamo la liphatsa tsa lefutso tse hlalositsoeng hantle, tse fapaneng ka ho hlaka ho tloha libakeng tsa Tropea le tsa Montoro tseo hangata li fosang ka tsona. E le ho fana ka tlhaloso ea li-bulbs, tse atisang ho sebelisoa bakeng sa tšebeliso e ncha, ho ile ha hlahlobjoa lintho tse tiileng tse qhibilihang le ho soeufala, ho bontša monate o phahameng ho ARO mabapi le libaka tse peli tse boletsoeng ka holimo. Ka kakaretso, thuto ea hajoale e na le thuso bakeng sa ho phahamisa boleng ba ARO nakong e tlang, e ka khothaletsoang ka lileibole tsa boleng tse ka tlatsetsang ho fokotsa bomenemene khoebong le ho ntlafatsa lekeno la lihoai tse nyane.
Selelekela
Mofuta oa Allium o kenyelletsa mefuta e ka bang 750 [1], eo har'a eona onion (Allium cepa L., 2n = 2x =16) e leng e 'ngoe ea tse atileng ka ho fetisisa. A. cepa e na le potoloho ea lilemo tse peli le boitšoaro bo fetang ba pelehi. Matsatsing ana, tlhahiso ea onion lefatšeng ka bophara (97.9 Mt) e e etsa sejalo sa bobeli sa bohlokoa sa meroho kamora tamati [2]. Ho tloha mehleng ea khale, li-bulbs tsa onion li 'nile tsa sebelisoa e le lijo le litlhare tsa setso. Ha e le hantle, Baegepeta ba boholo-holo ba ne ba se ba ntse ba tlaleha mekhoa e mengata ea phekolo e thehiloeng tšebelisong ea konofolo le eiee ho papirase ea bongaka ea 1550 BC, Codex Ebers [3].
Moroho ona o feto-fetohang le o phetseng hantle o jeoa o le tala, o le foreshe, kapa e le sehlahisoa se fetisitsoeng, 'me o sebelisoa ho ntlafatsa tatso ea lijana tse ngata. Liphuputso tse 'maloa tsa morao tjena li bolela hore tšebeliso ea onion e ka fokotsa kotsi ea mafu a pelo [4,5], botenya [6], lefu la tsoekere [7], le mefuta e sa tšoaneng ea kankere [8-10]. Lintho tse molemo tsa bophelo bo botle ba onion hangata li bakoa ke maemo a phahameng a lihlopha tse peli tsa metsoako ea phepo e nepahetseng: flavonoids le alk(en)yl cysteine sulphoxides (ACSOs). Sehlopha sa pele se kenyelletsa flavonols le anthocyanins. Quercetin ke flavonol e ka sehloohong e bonoang, e tsejoang ka thepa ea eona e matla ea antioxidant le e khahlanong le ho ruruha ho chefo le ho tlama li-ion tsa tšepe. [11]; athe li-anthocyanins li fana ka 'mala o mofubelu / o pherese mefuteng e meng ea onion. Ha e le li-ACSO, tse ngata ka ho fetisisa ke isoalliin [(+)-trans-S-1-propenyl-L-cysteine sulfoxide] [12], amino acid e sa fetoheng le e seng proteinogenic sulfur amino acid e bolokiloeng liseleng, e ikarabellang ka tsela e sa tobang bakeng sa monko o monate le tatso ea eiee. [13]. Ha lisele li senyeha, isoalliin e petsoha ke enzyme alliinase ho hlahisa letoto la metsoako e sa tsitsang (pyruvate, ammonia, thiosulphonates le propanethial S-oxide) e bakang ho taboha le ho baka monko o sa thabiseng (pungency). [14]. The onion pungency hangata e lekanngoa e le palo, ka gram ea boima bo bocha, ea pyruvic acid e hlahisoang ke hydrolysis. [15,16].
Linaheng tsa Mediterranean beisine, sisintsweng e le e 'ngoe ea bobeli mefuta e fapaneng litsi tsa A. cepa [17,18], bulbs ea onion e na le mefuta e mengata e fapaneng ea sebōpeho, boholo, 'mala, ntho e omeletseng le pungency [19-22]. Ho feta moo, manyolo a thehiloeng ho sebabole, mekhoa ea temo, mofuta oa mobu, maemo a leholimo, le mofuta oa mofuta oa limela kapa limela tsa naha li ka susumetsa boleng ba tleloubu ka ho fana ka litekanyetso tse ikhethang tsa organoleptic le phepo e nepahetseng. [23-27]. Naheng ea Italy, ho sa tsotellehe ho fumaneha ha likokoana-hloko tsa onion ka bongata, ke mefuta e fokolang feela ea eiee e atisang ho etsoa lithuto tsa mahlale 'me e tšoauoa hantle. [28,29].
Sebopeho se hlakileng sa liphatsa tsa lefutso le tsa phenotypic tsa mefuta-futa ea agro-biodiversity ke ea bohlokoa ho netefatsa paballo e nepahetseng ea mehloli ea liphatsa tsa lefutso tsa semela le ho khothaletsa ts'ebeliso ea mefuta e ikhethileng ea ketane ea boleng. [30-32]. Hangata ho khethiloe matšoao a tatellano a bonolo (SSR) bakeng sa 'mapa [33-35], khatiso ea menoana ea DNA le khethollo ea mefuta [36-38], le khakanyo e tšepahalang ea phapang ea liphatsa tsa lefutso ka hare le har'a libaka tsa naha [39-42], kaha li na le li-locus specific, li-allelic tse ngata, li futsitsoe ka bonngoe, li ka ikatisa haholo, 'me li loketse genotyping e ikemetseng.
Thutong ea hona joale, re tsepamisitse maikutlo a rona sebakeng sa setso sa Apulian, "Acquaviva red onion" (ARO), e lengoang ho latela mekhoa ea temo ea manyolo sebakeng se senyenyane sa toropo ea Acquaviva delle Fonti, profinseng ea Bari. (Apulia, Italy Boroa). Li-bulbs tsa landrace ena li kholo 'me li bataletse ebile li na le mebala e khubelu' me li sebelisoa haholo lijong tsa lehae. Leha ARO e fumane letšoao la boleng la "Slow Food Presidium", tlhahiso ea eona e ka khothaletsoa le ho sireletsoa ke matšoao a boleng a European Union joalo ka ponts'o ea sebaka se sirelelitsoeng (PGI) le lebitso le sirelelitsoeng la tlhaho (POD), kaha tsena li ka kenya letsoho ho fokotsa bomenemene khoebong le ho ntlafatsa chelete ea lihoai tse nyane. Mona, matšoao a limolek'hule a SSR a ne a sebelisoa e le lisebelisoa tse matla tsa ho lekola phapang ea liphatsa tsa lefutso har'a baahi ba ARO le ho khetholla sebaka sena ho tsoa libakeng tse ling tse peli tsa eiee e khubelu ea Italy. Ho feta moo, re hakantse lintho tse thata le tse qhibilihang ka har'a metsi ho lekola tatso ea ARO mabapi le tlhoko ea 'maraka.
Results
Ho thehoa ha Acquaviva Red Onion Germplasm Collection le Morphological Characterization
Peo ea baahi ba 13 ba ARO landrace, e fanoeng ke lihoai moralong oa projeke ea Lebatooa la BiodiverSO Apulia e sebelisitsoe ho theha pokello ea likokoana-hloko tsa ARO.
Litlhaloso tsa morphological, tse amanang le tleloubu, letlalo le nama li ile tsa bokelloa holim'a ARO germplasm le holim'a libaka tse tharo tsa onion, tse peli tsa "Tropea red onion" (TRO) landrace le e 'ngoe ea "Montoro copper onion" (MCO) landrace (Setšoantšo 1). Li-bulbs tsohle tsa ARO li ne li bataletse 'me li ne li khetholloa ke letlalo le khubelu la ka ntle le nama e nang le mebala e sa tšoaneng ea bofubelu. Ka lehlakoreng le leng, nama ea li-TRO bulbs e ne e le khubelu ka ho feletseng, athe nama ea bulbs ea MCO e ne e se na 'mala o mobe (Letlapa la S1). Tlhahlobo ea biochemical e lumelletsoe ho lekola litaba tse qhibilihang ka har'a metsi le pungency. Joalo ka ha ho tlalehiloe ho Tafole 1, kakaretso ea boleng ba lintho tse tiileng tse qhibilihang tsa bulbs ho baahi ba ARO e ne e le 7.60, 'me e le ho tloha ho 6.00 (ARO12) ho isa ho 9.50° Brix (ARO11 le ARO13). Boleng bona bo ne bo phahame ho feta bo hakantsoeng bakeng sa libaka tsa naha tsa TRO le MCO (4.25 le 6.00° Brix, ka ho latellana).
Tafole 1. Solid Soluble Content le Pungency Values Assessed in "Acquaviva Red Onion" (ARO), "Tropea Red Onion" (TRO), le "Montoro Copper Onion" (MCO) Populations *.
CODE | Soluble Solid Content (Brix) | Pungency (pmolg-1 FW) | ||
E bolelang | CV y (%) | E bolelang | CV y (%) | |
ARO1 | 6.25D * | 5.65 | 5.84 ab * | 23.78 |
ARO2 | 7.25 DC | 4.87 | 6.51 ho | 22.98 |
ARO3 | 7.50BCD | 9.42 | 5.28ab | 22.88 |
ARO4 | 7.50BCD | 0.00 | 6.97 ho | 3.74 |
ARO 5 | 7.50BCD | 0.00 | 6.80 ho | 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 ho | 3.49 |
ARO9 | 8.25 ABC | 4.28 | 6.84 ho | 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 ho | 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 |
* Meelelo e nang le litlhaku tse tšoanang ka litlhaku tse kholo kapa tse nyane ha li fapane ka lipalo ho 0.01P kapa 0.05P, ka ho latellana (Teko ea SNK). y Coefficient ea phapang.
Theko e tloaelehileng ea ARO pungency, e hlahlobiloeng ka pyruvic acid content, e ne e le 6.00, e tloha ho 4.51 pmol g.-1 FW (ARO6) ho isa ho 7.04 (ARO8). Boleng bona bo ne bo le holimo ho feta bo hakanyetsoang ho TRO le MCO landraces (3.54 pmol g-1 FW le 4.18 pmol g-1 FW, ka ho latellana).
SSR Polymorphism le Likamano tsa Liphatsa tsa lefutso har'a Accessions
Phuputsong ea hajoale, metsoako e 11 ho e 37 e lekiloeng ea SSR e fane ka polymorphisms ea locus e le 'ngoe, ke hore, e fana ka boholo ba lihlahisoa tse peli tsa amplification ho motho a le mong. Ka kakaretso, li-allele tsa 55 li ile tsa fumanoa ho batho ba 320 ba nang le li-alleles tse ngata ho tloha ho 2 (ACM147 le ACM 504) ho ea ho 11 (ACM132) le boleng bo bolelang ba 5 alleles (Letlapa). 2). Ho batho ka bomong, palo ea alleles (Na) e ne e tloha ho 1.94 (ACM147 le ACM504) ho ea ho 5.38 (ACM132), athe palo e sebetsang ea alleles (Ne) e ne e tloha 1.41 (ACM152) ho ea 2.82 (ACM449). Liphapang pakeng tsa boleng ba Na le Ne li ne li bakoa ke ho ba teng ha alleles tse nang le maqhubu a tlaase ho baahi le ho ata ha li-allele tse seng kae feela. Ntho e phahameng ka ho fetisisa e hlokometsoeng ea heterozygosity (Ho) e ile ea totobatsoa bakeng sa ACM138 le ACM449 (0.62), athe e tlaase ka ho fetisisa e ne e amahanngoa le ACM152 (0.25). Ho lebeletsoe heterozygosity (He), e lumellanang le tebello ea theory ho baahi ba panmictic, ho tloha ho 0.37 (ACM504) ho ea 0.61 (ACM132, ACM138, le ACM449). Wright's fixation index (Fis), e bonts'itseng boleng bo haufi le lefela (karolelano ea 0.05) bakeng sa matšoao ohle, e bonts'ang boleng bo tšoanang lipakeng tsa maemo a heterozygosity a hlokometsoeng le a lebelletsoeng, joalo ka ha ho lebelletsoe bakeng sa mefuta e fetang. Ho sebetsa hantle ha letšoao la motho ka mong la SSR ka ho hatisa menoana ea liphatsa tsa lefutso ho ne ho hakanngoa ke index ea polymorphic information content (PIC), e nang le boleng ba 0.48 le ho tloha 0.33 (ACM504) ho 0.67 (ACM132). Lenane le leng la katleho, Shannon's Information Index (I) e bonts'itse boleng ba boleng ba 0.84, 'me ho nahanoa hore boleng bo tloha ho 0.45 (ACM152) ho isa ho 1.20 (ACM132).
Tafole 2. Likarolo tsa Polymorphism tsa Lits'oants'o tsa 11 tsa SSR tse Sebelisitsoeng ho Hakanya Phapang ea Liphatsa tsa lefutso ho ARO, TRO, le MCO Populations. Kakaretso ea Nomoro ea Alleles (Na), Band Size Range, le Polymorphic Information Content (PIC) Index Ho Sheba Kakaretso ea Sehlopha sa Batho ba 320 ba Genotype Thutong ena. Nomoro ea Alleles (Na), palo ea Alleles e Atlehang (Ne), E Sheterozygosity e Bonoeng (Ho), Ho Lebeletsoe Heterozygosity (He), Fixation Index (Fis), le Index ea Boitsebiso ea Shannon (I) e bua ka Mean Values e Baloang ho tsoa ho Baahi ba 16, E ’ngoe le e ’ngoe e Kopantsoe ke Batho ba 20.
Locus. | Kakaretso Na | Size Range (bp) | PIC | E bolelang | |||||
Na | Ne | Ho | He | I | Fis | ||||
EA-91-ACM | 4 | 189-205 | 0.40 | 2.63 | 1.72 | 0.38 | 0.39 | 0.66 | 0.04 |
EA-101-ACM | 4 | 229-241 | 0.52 | 2.94 | 2.37 | 0.53 | 0.56 | 0.92 | 0.06 |
EA-132-ACM | 11 | 186-248 | 0.67 | 5.38 | 2.78 | 0.55 | 0.61 | 1.20 | 0.09 |
EA-138-ACM | 5 | 242-272 | 0.66 | 3.69 | 2.82 | 0.62 | 0.61 | 1.09 | -0.02 |
EA-147-ACM | 2 | 264-266 | 0.37 | 1.94 | 1.83 | 0.44 | 0.44 | 0.62 | -0.01 |
EA-152-ACM | 4 | 228-244 | 0.25 | 2.38 | 1.41 | 0.25 | 0.27 | 0.45 | 0.07 |
EA-235-ACM | 4 | 286-298 | 0.41 | 2.81 | 1.77 | 0.44 | 0.41 | 0.72 | -0.06 |
EA-446-ACM | 6 | 108-120 | 0.56 | 3.50 | 2.48 | 0.49 | 0.58 | 1.01 | 0.16 |
EA-449-ACM | 8 | 120-140 | 0.66 | 4.88 | 2.82 | 0.62 | 0.61 | 1.18 | -0.03 |
EA-463-ACM | 5 | 202-210 | 0.47 | 3.38 | 1.95 | 0.46 | 0.48 | 0.83 | 0.05 |
EA-504-ACM | 2 | 188-192 | 0.33 | 1.94 | 1.64 | 0.30 | 0.37 | 0.54 | 0.20 |
E bolelang | 5 | 0.48 | 3.22 | 2.15 | 0.46 | 0.48 | 0.84 | 0.05 |
Har'a baahi, ARO3, ARO6, ARO8, ARO10, TRO1, le MCO li bonts'itse boemo bo phahameng ba phapang ea liphatsa tsa lefutso (Ho > 0.5), athe mefuta e tlaase ka ho fetisisa e ile ea hlokomeloa ho baahi ba ARO7 (Ho = 0.27) (Tlaleho ea Tlatsetso S2). Ka kakaretso, likhetho tsohle li bonts'itse Fis boleng bo haufi le zero (Fis ho bolela boleng = 0.054), joalo ka ha ho lebeletsoe tlas'a maemo a sa tloaelehang a ho nyalana.
Tlhahlobo ea Phapang ea Limolek'hule le Sebopeho sa Liphatsa tsa lefutso
Karohano ea maemo a fapaneng ea liphatsa tsa lefutso har'a le ka har'a baahi e ile ea etsoa computed ke AMOVA. Liphetho li bonts'itse karolo e kholo ea phapano ea liphatsa tsa lefutso ka har'a baahi (87%). Phapang pakeng tsa baahi, 13%, e ne e le ea bohlokoa haholo (P <0.001) (Tafole 3). Litekanyetso tse tsamaeang ka bobeli tsa paramethara ea Fpt, e tšoanang le index ea Wright's Fst fixation, ho tloha ho 0.002 (ARO2/ARO10) ho isa ho 0.468 (ARO7/TRO2), e ne e le ea bohlokoa (P <0.05), ntle le dipapiso tse robong ka bobedi (Tlaleho ya Tlatsetso S3).
Tafole 3. Tlhahlobo ea Phapang ea Limolek'hule ea 320 Genotypes ho tloha ho 16 Baahi ba allium cepa L.
mohloli o moholo | df | Kakaretso ea Libaka | Phapang Khakanyo | Phapang (%) | Fpt | P |
Har'a baahi | 15 | 458.63 | 1.16 | 13% | ||
Ka har'a baahi | 304 | 2272.99 | 7.50 | 87% | 0.134 | 0.001 |
Kakaretso | 319 | 2731.62 | 8.66 |
Patlisiso ea sebopeho sa liphatsa tsa lefutso ho A. cepa pokello ea genotyped thutong ena e entsoe ka tlhahlobo e kopaneng ea moetso oa lihlopha tse kentsoeng tšebetsong ho STRUCTURE ea software. Mokhoa oa Evanno AK o khothalelitse karohano ka lihlopha tse peli (K = 2) e le eona e rutang haholo bakeng sa rona. setatata,le the E 'ngoe phahameng ka ho fetisisa peak ho K = 5 (Sopplementaiv Rgure S1). A bakeng sa K = 2, ahpoplitloaelo weseleigned ho onle eaf lihlopha tse peli le khoeli ea rnernbertoip (q) > 0.7. Joalo ka shown ka Figure 2a, sehlopha sa pele (se bitsoang S1) se ne se kenyelletsa MCO le baahi bohle ba ARO, athe sehlopha sa S2 se ne se arola lihlopha tse peli tsa TRO. Ho K = 5, ho fana ka tlhaloso e tebileng ea dataset (Setšoantšo 2b), 75% ea likhetho li ile tsa abeloa ho e 'ngoe ea lihlopha tse hlano. Karohano pakeng tsa ARO (S1) le TRO (S2) e tiisitsoe, le hoja batho ba bang ba ARO ba ne ba kopantsoe (q <0.7) kapa ba arotsoe ka lihlopha tse peli tse ncha tsa S3 le S4 (ARO7 le ARO12, ka ho latellana). Hoa thahasellisa hore mofuta oa khoebo oa MCO o thehile sehlopha se ikhethileng (S5) se arohaneng le onion e khubelu ea Apulian.
Likamano tsa liphatsa tsa lefutso har'a Baahi
SSR polymorphism e lumelloa ho hula dendrogram ea mefuta-futa ea liphatsa tsa lefutso 'me liphello tsa tlhahlobo ea phylogenetic li bontšoa ho Figure. 3a. Mona, pokello ea germplasm e arotsoe ka lihlopha tse hlano tse tšehetsoeng ka matla ke litekanyetso tsa bootstrap. Baahi ba ARO7 le ARO12 ba ile ba aroloa hang-hang ho tsoa ho baahi ba setseng mme ba theha lihlopha tse peli tse ikhethileng. Sehlopha sa boraro se kenyelelitse batho ba babeli ba khoebo ba TRO, khabareng sebaka sa bone se arola MCO ho tsoa ho batho ba leshome le motso o mong ba ARO. Kamano ea liphatsa tsa lefutso e hlahang har'a baahi e ile ea boela ea etsoa lipatlisiso ka principal coordinate analysis (PCoA) (Figure 3b). Joalo ka ha ho totobalitsoe pejana, baahi ba ARO ba ne ba hlophisoa ka thata, ntle le ARO12 le ARO7, tse hlahileng maemong a ka thoko ho morero oa PCoA. Li-TRO tse peli le batho ba MCO ba ne ba hasane karolong e ka tlase-le letona la morero.
Setšoantšo sa 3. Phapang ea liphatsa tsa lefutso har'a 16 A. cepa palo ea batho ba tšoailoeng thutong ena, ho ipapisitsoe le boemo ba bona ba SSR. (a) UPGMA dendrogram ea sebaka sa liphatsa tsa lefutso. Litekanyetso tsa tšehetso ea Bootstrap> 50 li bonts'itsoe ka holim'a li-node tse lumellanang; (b) principal component analysis (PCoA). Sehlopha se pota-potiloe ka bofubelu bo lumellanang ka ho feletseng le sehlopha se hlahisoang ke tlhahlobo ea phylogenetic 'me se thehiloe ke 11 ARO accessions.
Puisano
Ka har'a palo e kholo ea mefuta-futa ea li-agro-biodiversity tse lengoang ka tloaelo Italy Boroa, mabala a onion a emela lihlahisoa tse hlokang ho sireletsoa kotsing ea khoholeho ea liphatsa tsa lefutso le tšokelo ea ho nkeloa sebaka ke mefuta ea sejoale-joale. Ka moralo oa projeke ea tikoloho ea BiodiverSO, e reretsoeng ho bokella, ho tšoaea, ho khothaletsa le ho sireletsa mehloli ea liphatsa tsa lefutso tsa sebaka sa Apulia tse amanang ka matla le lefa la lehae, re thehile pokello ea peo ea baahi ba 13 ba ARO landrace. Re tlalehile tlhahlobo ea pele ea phapang ea ARO ho latela li-polymorphisms tsa DNA le li-parameter tse peli tsa biochemical, li-soluble solid le pyruvic acid content, tse amanang le litšobotsi tsa tatso le tsa bohlokoa bakeng sa ho amoheloa ha lihlahisoa tse ncha tse sa phehoang. Ntle le moo, lintlha tse mabapi le ARO landrace li ne li bapisoa le tse bokelletsoeng libakeng tse ling tse peli tse nang le 'mala oa onion tseo hangata li fosang ka tsona.
Litlhahlobo tsa lik'hemik'hale li bonts'itse monate oa batho ba 13 ba ARO, ba amanang le lintho tse ngata tse qhibilihang ka har'a metsi le bohloeki bo bohareng, ho latela litataiso tsa indasteri ea onion e monate. [31]. Li-bulbs tsa ARO li ne li le monate ho feta tsa libaka tsa naha tsa TRO le MCO, 'me li ne li bontša bohale bo phahameng hanyane. Leha ho le joalo, monate oa eiee o bakoa ke ho leka-lekana pakeng tsa tsoekere le pungency, kahoo tšobotsi ena e ka ba molemo ho tšehetsa khetho ea genotypes ea boleng, hangata e etsoang ke lihoai tse itšetlehileng feela ka morpholoji.
Lits'oants'o tsa SSR li netefalitsoe e le sesebelisoa sa bohlokoa sa ho khetholla mefuta ea liphatsa tsa lefutso, leha e bokelloa ka har'a sebaka se moqotetsane se ntseng se hola joalo ka toropo ea Acquaviva delle Fonti. Lits'oants'o tse khethiloeng li bonts'itse palo e phahameng ea li-allele ho feta matšoao a tlalehiloeng pele [43] 'me [44], empa e le tlase ho feta matšoao a tlalehiloeng ke [45]. Ho feta moo, 50% ea sete ea matšoao a rona e bonts'itse boleng ba index ea PIC e kholo ho feta 0.5, e bonts'a hore e loketse ho khetholla batho ba pokellong, joalo ka ha ho khothalelitsoe ke [46]. Tekolo ea mefuta-futa ka har'a baahi e senotse litekanyetso tse tšoanang lipakeng tsa Ho le He, e leng se fellang ka hore Fi e be tlases litekanyetso. Sena se lumellana le mokhoa oa ho tsoa oa A. cepa, e tshwerweng haholo ke inbreeding depression [47]. Kakaretso ea Fis boleng bo baloang ho palo ea onion e nahanoang thutong ena (0.054) e ne e le tlase ho feta e tlalehiloeng pele ke [45] (0.22) mme e batla e ts'oana le e fumanoeng ke [31] (0.08) le [48] (0.00) ea ileng a lekola mefuta-futa ea liphatsa tsa lefutso libakeng tsa onion ho tloha leboea-bophirima ho Spain le Niger, ka ho latellana. Maemo a hlokomelehang a heterozygosity ho baahi ba ARO a tiisa taba ea hore Apulia e emela setsi sa mefuta-futa sa mefuta e mengata ea lirapa. [32, 42, 49-51].
AMOVA e hlakisitse hore phapang e ngata ea limolek'hule pokellong ea liphatsa tsa lefutso thutong ena e ka har'a batho. Leha ho le joalo, phapang ea bohlokoa ea liphatsa tsa lefutso har'a baahi (FPT values) e senotse ketsahalo ea stratification ea liphatsa tsa lefutso. Ha e le hantle, le hoja liphetho tsa rona li bontšitse ho ba teng ha liphatsa tsa lefutso ho batho ba bangata ba ARO, ho theha sehlopha se hlalositsoeng hantle, baahi ba ARO7 le ARO12 ba bontšitse boitsebiso bo hlakileng ba lefutso. Sephetho sena se ka bakoa ke tšimoloho e fapaneng ea peo e sebelisoang ke lihoai tse peli tseo baahi ba neng ba bokelloa ho tsona. Ho feta moo, ho latela liphetho tse fumanoeng, sebaka sa ARO se ka nkoa se fapane ka ho hlaka boemong ba liphatsa tsa lefutso ho tloha ho TRO le MCO landraces. Phuputsong ea morao tjena, [29] e ile ea lekola mefuta-futa ea liphatsa tsa lefutso tsa libaka tse 'maloa tsa eiee ea Italy ho kenyelletsa "Acquaviva," "Tropea," le "Montoro." Le hoja bangoli ba sebelisitse matšoao a SNP ho hlahloba mefuta-futa ea liphatsa tsa lefutso tsa pokello ea onion e pharaletseng, genotyping e ne e sa khone ho khetholla "Acquaviva" ho tloha "Tropea" le "Montoro" eiee. Mohlomong, phapang ena e bakoa ke boleng bo tlase ba PIC bo fumanoeng (0.292), bo fanang ka maikutlo a boitsebiso bo itekanetseng bo akaretsang ba sebaka se ntseng se hlahlojoa joalo ka ha ho boletsoe ke [29]. Ho feta moo, molemong oa ho batlisisa boteng ba sub-structure sehlopheng sa bona sa Italy, ho ka be ho bile molemo ho sekaseka genotypes ea Italy ka thoko ho tse ling kaofela tsa pokello. Mohlomong e ka be e ile ea lumella ho bona ka mahlo a kelello mohlala oa mefuta-futa ea liphatsa tsa lefutso e amanang le stratification ea libaka kapa mekhoa e tlas'a khetho e matla.
Qetellong, phuputso ea hona joale e emela tlaleho e pharaletseng ea sebaka sa onion se amanang le lefa la setso sa lehae le bohlokoa ba moruo ho lihoai. Liphello tsa rona li totobatsa hore, ntle le mekhelo e seng mekae, ARO e khetholloa ke liphatsa tsa lefutso tse hlalositsoeng hantle, tse lokelang ho sireletsoa kotsing ea khoholeho ea liphatsa tsa lefutso. Ka hona, ho thehoa ha pokello ea baemeli ba mohloli ona oa bohlokoa oa mefuta-futa ea liphatsa tsa lefutso e bile habohlokoa. Qetellong, sebopeho sa lefutso le phenotypic sa ARO se ka ba molemo ho fumana matšoao a boleng ho tsoa European Union.
Lisebelisoa le mekhoa
Pokello ea Likokoana-hloko, Semela sa Semela, le Phallo ea DNA
Sehlopha sa baahi ba 13 ba sebaka sa ARO se ile sa fumanoa ka har'a moralo oa morero oa Apulia Region (BiodiverSO: https://www.biodiversitapuglia.it/), ka letoto la mesebetsi e etsoang "Acquaviva delle Fonti", toropo e nyenyane ea Apulian Profinseng ea Bari, Italy. Libaka tsa pokello ea sebaka se seng le se seng li ile tsa etsoa 'mapa ka Geographic Information System (GIS) 'me tsa tlalehoa ho Lethathamo 4. Ho phaella moo, batho ba babeli ba tsoang TRO landrace le baahi ba le bang ba tsoang MCO landrace ba kenyelelitsoe thutong ea hona joale 'me ba sebelisoa e le litšupiso. Lihlahisoa tsohle tsa semela li ne li lengoa maemong a tšoanang a tikoloho polasing ea liteko "P Martucci" ea Univesithi ea Bari (41° 1'22.08″ N, 16°54'25.95″ E), tlas'a lesaka la tšireletso ho qoba ho tsamaisa peō lipakeng tsa tsona. palo ea batho le ho netefatsa phallo ea batho ka har'a baahi ka lintsintsi (Lucilia Cesare). Baahi ba 16 ba ne ba khetholloa ka litšobotsi tse amanang le boholo ba tleloubu le sebopeho le 'mala oa letlalo le nama (Letlapa la S1). Ho feta moo, tlhahlobo e tiileng ea lintho tse qhibilihang e entsoe ho sebelisoa refractometer e tšoaroang ka letsoho, 'me pungency e ile ea lekanngoa ka lisampole tsa lero la onion ho eketsa 2,4-dinitrophenyl hydrazine (0.125% v/v ka 2N ea HCl) le ho hlahloba monyetla ho 420 nm, joalokaha ho tlalehiloe ke [31]. Teko ea Duncan ea mefuta e mengata le tlhahlobo ea SNK li ile tsa etsoa ho fumana boteng ba liphapang tse kholo.
Tafole 4. Lethathamo la Baahi ba Bokelitsoeng le ba Fetotsoeng Thutong ena. Bakeng sa Baahi ba Mong le Bong, Khoutu ea Boitsebiso, Lebitso la Sebaka, GPS Coordinate, le Banka ea Gene e Bolokang Peo li tlalehoa.
Code | lebitso | Lihokela tsa GPS | Gene Bank y |
ARO1 | Cipolla rossa ea Acquaviva | 40°54’21.708″ N 16°49’1.631” E | Di.SSPA |
ARO2 | Cipolla rossa ea Acquaviva | 40°53’14.28″ N 16°48’56.879” E | Di.SSPA |
ARO3 | Cipolla rossa ea Acquaviva | 40°54’11.304″ N 16°49’13.079” E | Di.SSPA |
ARO4 | Cipolla rossa ea Acquaviva | 40°54’3.348″ N 16°40’27.011” E | Di.SSPA |
ARO5 | Cipolla rossa ea Acquaviva | 40°51’59.76″ N 16°53’0.527” E | Di.SSPA |
ARO6 | Cipolla rossa ea Acquaviva | 40°52’48.72″ N 16°49’43.247” E | Di.SSPA |
ARO7 | Cipolla rossa ea Acquaviva | 40°53’13.47″ N 16°50’23.783” E | Di.SSPA |
ARO8 | Cipolla rossa ea Acquaviva | 40°53’18.816″ N 16°49’33.888” E | Di.SSPA |
ARO9 | Cipolla rossa ea Acquaviva | 40°54'51.372 ″ N 16°49'3.504" E | Di.SSPA |
ARO10 | Cipolla rossa ea Acquaviva | 40°54’1.188″ N 16°49’24.311” E | Di.SSPA |
ARO11 | Cipolla rossa ea Acquaviva | 40°52'49.8 ″ N 16°49'48.575" E | Di.SSPA |
ARO12 | Cipolla rossa ea Acquaviva | 40°52’38.892″ N 16°49’28.379” E | Di.SSPA |
ARO13 | Cipolla rossa ea 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, Lefapha la Mobu, Semela le Lijo tsa Saense, Univesithi ea Bari. |
Lisebelisoa tsa makhasi a 20 genotypes ka baahi li ile tsa nkoa 'me tsa bolokoa ho -80 °C ho fihlela li sebelisoa. Bakeng sa mefuta e mengata ea polysaccharide, joalo ka A. cepa, mehato ea pele ea ho tlosa polysaccharide e bohlokoa ho fumana DNA ea boleng bo botle, ka hona litlhatso tsa pele ho STE buffer (0.25 M sucrose, 0.03 M Tris, 0.05 M EDTA) li entsoe joalo ka ha ho hlalositsoe ke [52]. Kakaretso ea DNA e ntšitsoe ho latela mokhoa oa CTAB [53] 'me qetellong e ile ea hlahlojoa boleng le mahloriso ke Nano Drop 2000 UV-vis spectrophotometer (ThermoScientific, Waltham, MA, USA) le 0.8% ea agarose gel electrophoresis.
Tlhahlobo ea SSR
16 EST-SSR metsoako ea mantlha e ntlafalitsoeng ke [54] le ho lekoa pele lithutong tsa mefuta-futa ea liphatsa tsa lefutso ka [43] 'me [44] le 21 genomic SSR [45-55] li ile tsa hlahlojoa ho lekola ho tšoaneleha ha tsona (Tlaleho ea Tlatsetso ea S4). Genotyping e ne e etsoa ho sebelisoa mokhoa oa ho tšoaea oa moruo oa fluorescent moo mohatla oa M13 o kenyellelitsoeng ho e 'ngoe le e' ngoe ea pele ea SSR. [56]. Li-mixx tsa PCR li entsoe ka 20 gL reaction e nang le: 50 ng ea kakaretso ea DNA, 0.2 mM ea dNTP mix, 1X ea PCR reaction buffer, 0.8 U ea DreamTaq DNA polymerase (Thermo Scientific, Waltham, MA, USA), 0.16 gM ea reverse primer , 0.032 gM ea primer ea pele e atolositsoeng ka tatellano ea M13 (5′-TGTAAAACGACGGCCAGT-3′), le 0.08 gM ea M13 ea bokahohle e ngotsoeng ka lidae tsa fluorescent tsa FAM kapa tsa NED (Sigma-Aldrich, St. Louis, MO, USA). Karabelo ea PCR e entsoe ka SimpliAmp (Applied Biosystems, CA, USA) thermocycler e nang le maemo a latelang bakeng sa boholo ba lipara tsa pele: 94 °C bakeng sa metsotso e 5, lipotoloho tse 40 ho 94 °C bakeng sa 30 s, 58 °C. bakeng sa 45 s le 72 °C bakeng sa 45 s le bolelele ba ho qetela ho 72 °C bakeng sa 5 mets. Ha e le ACM446 le ACM449, PCR ea touchdown e ile ea sebelisoa ka ho kenngoa ha 60 ° C ho 55 ° C ho feta lipotoloho tse 10, lipotoloho tse 30 ho 55 ° C, tse lateloang ke katoloso ea ho qetela ea 5 min ho 72 ° C. Lihlahisoa tsa PCR li ile tsa kenngoa ka har'a poleiti ea seliba sa 96 'me e kopantsoe le 14 gL ea Hi-Di Formamide (Life Technologies, Carlsbad, CA, USA) le 0.5 gL GeneScan 500 ROX Size Standard (Life Technologies, Carlsbad, CA, USA). Li-amplikhone li ile tsa rarolloa ka mochini oa tatellano oa li-capillary oa ABI PRISM 3100 Avant Genetic Analyzer (Life Technologies, Carlsbad, CA, USA), moo li-allele li ileng tsa fuoa lintlha e le tse laolang 'me tsa abeloa ka ho sebelisa GeneMapper Software Version 3.7.
Lisebelisoa tsa GenAlEx 6.5 [57] le Cervus 3.0.7 [58] li ne li sebelisetsoa ho hakanya palo ea alleles (Na), palo ea alleles e sebetsang hantle (Ne), heterozygosity (Ho), heterozygosity e lebelletsoeng (He), polymorphic information content (PIC), index ea tlhahisoleseling ea Shannon (I), le fixation index (Fis). ) bakeng sa sebaka se seng le se seng sa SSR.
Tekolo ea Phapano ea Liphatsa tsa lefutso
Karohano ea maemo a fapaneng ea liphatsa tsa lefutso har'a le ka har'a baahi ba onion e ile ea hlahlojoa ke GenAlEx 6.5 [57] ka tlhahlobo ea phapang ea limolek'hule (AMOVA) le 999 bootstrapping ho hlahloba bohlokoa. Ho feta moo, software ea GenAlEx 6.5 e ile ea sebelisoa ho hakanya phapang e teng har'a sechaba ka seng ka ho etsa komporo ea karolelano ea Ho, He, le Fis holim'a libaka tsohle tsa SSR.
Sebopeho sa baahi se ile sa nkoa ke mokhoa oa Bayesian-based clustering algorithm e sebelisoang ho STRUCTURE v.2.3.4 software [59]. Seteishene sa data se ne se tsamaisoa ka lihlopha tse 'maloa tse inahaneloang (K), ho tloha ho 1 ho isa ho 10, ho beha li-run tse leshome tse ikemetseng ho boleng bo bong le bo bong ba K. Bakeng sa lebelo le leng le le leng, sepheo sa ho netefatsa ho lumellana ha liphetho, nako ea ho chesoa ea pele ea 100,000 le liphetolelo tse 100,000 tsa Markov Chain Monte Carlo (MCMC) li entsoe tlas'a mohlala oa motsoako le maqhubu a ikemetseng har'a batho. Boleng bo ka bang teng ba K bo ne bo ikemiselitse ho kenya tšebetsong mokhoa oa AK, o hlalosoang ke [60], lenaneong la marang-rang la STRUCTURE HARVESTER [61]. Batho ka bomong ba ne ba abetsoe sehlopha se itseng ha palo ea litho tsa eona (q-value) e ne e le holimo ho 0.7, ho seng joalo e ne e nkoa e le leloko le tsoakiloeng.
Tlhahlobo e ka sehloohong ea ho hokahanya e entsoe e le ho bona ka mahlo a kelello mekhoa ea kamano ea liphatsa tsa lefutso har'a likhetho tse senotsoeng ke Nei's genetic distance matrix (Supplementary Table S5). Ho ipapisitsoe le maqhubu a liphatsa tsa lefutso, dendrogram ea sebaka sa liphatsa tsa lefutso e ile ea etsoa ho phethahatsa mokhoa oa sehlopha se se nang boima ba 'mele ka tlhahlobo ea lihlopha tsa lipalo tsa lipalo (UPGMA) ho software ea POPTREEW. [62]. Bootstrapping e ile ea sebelisoa ho lekola kholiseho ho sehlopha sa maemo a holimo, ho beha mohlala oa 100 oa sete ea data. Qetellong, software ea MEGA X [63] e ne e sebelisoa e le software ea ho taka lifate.
Lisebelisoa tsa Tlatsetso: Tse latelang li fumaneha inthaneteng ho http://www.mdpi.com/2223-7747/9/2/260/s1. Letlapa la S1: Sebopeho sa morphological ea li-bulbs tsa ARO, MCO, le TRO. Letlapa la S2: Heterozygosity le li-indices tsa fixation tse balloang bakeng sa landraces ea ARO le TRO le MCO landraces. Letlapa la S3: Maemo a mabeli a paramente ea Fpt. Lethathamo la S4: Lenane la li-SSR tse sebelisitsoeng thutong. Lethathamo la S5. Pairwise population matrix ea Nei genetic distance. Setšoantšo sa S1: Chate ea mela ea boleng ba K e fetohang le Delta K ea Evanno.
Menehelo ea Mongoli: CL le LR ba ile ba emola thuto mme ba rala teko; CL le PI li ile tsa etsa tlhahlobo ea li-molecular marker; ARM le VZ li ile tsa etsa liteko tsa tšimo; RM, SP, GR, le CL li ne li kentse letsoho tlhahlobong ea lintlha; RM le CL ba ngotse buka e ngotsoeng ka letsoho. Bangoli bohle ba balile mme ba lumellana le mofuta o hatisitsoeng oa buka e ngotsoeng ka letsoho.
Lithuso: Mosebetsi ona o tšehelitsoe ka lichelete ke morero oa Regional Apulian "Biodiversity of Apulian meroho mefuta" -Programma di Sviluppo Rurale per la Puglia 2014-2020. Misura 10—Sottomisura 10.2; fana ka CUP H92C15000270002, Italy.
lumela hore baa fokola: Liteboho li tsoa ho "Azienda Agricola Iannone Anna" le "Associazione produttori della vera cipolla rossa di Acquaviva" bakeng sa ho fana ka lisebelisoa tsa semela tse sebelisitsoeng tekong.
Likhohlano tsa Thahasello: Bangoli ba phatlalatsa hore ha ho na likhohlano tsa thahasello.
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