Development and utilization of new sequenced characterized amplified region markers specific for E genome of <italic>Thinopyrum

doi:10.1007/s11515-013-1268-9

Front Biol

2013, Vol. 8

Issue (4) : 451-459 https://doi.org/10.1007/s11515-013-1268-9

RESEARCH ARTICLE

Development and utilization of new sequenced characterized amplified region markers specific for E genome of Thinopyrum

Wenping GONG, Ling RAN, Guangrong LI, Jianping ZHOU, Cheng LIU(

), Zujun YANG

School of Life Science and Technology, University of Electronic Science and Technology of China, Chengdu 610054, China

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Abstract

Species containing E genome of Thinopyrum offered potential to increase the genetic variability and desirable characters for wheat improvement. However, E genome specific marker was rare. The objective of the present report was to develop and identify sequenced characterized amplified region (SCAR) markers that can be used in detecting E chromosome in wheat background for breeding purpose. Total 280 random amplified polymorphic DNA (RAPD) primers were amplified for seeking of E genome specific fragments by using the genomic DNA of Thinopyrum elongatum and wheat controls as templates. As a result, six RAPD fragments specific for E genome were found and cloned, and then were converted to SCAR markers. The usability of these markers was validated using a number of E-genome-containing species and wheat as controls. These markers were subsequently located on E chromosomes using specific PCR and fluorescence in situ hybridization (FISH). SCAR markers developed in this research could be used in molecular marker assisted selection of wheat breeding with Thinopyrum chromatin introgressions.

Keywords Thinopyrum Trititrigia E genome SCAR markers FISH

Corresponding Author(s): LIU Cheng,Email:liucheng@uestc.edu.cn

Issue Date: 01 August 2013

Cite this article:

Wenping GONG,Ling RAN,Guangrong LI, et al. Development and utilization of new sequenced characterized amplified region markers specific for E genome of Thinopyrum[J]. Front Biol, 2013, 8(4): 451-459.

URL:

https://academic.hep.com.cn/fib/EN/10.1007/s11515-013-1268-9
https://academic.hep.com.cn/fib/EN/Y2013/V8/I4/451

Species	Chromosome number	Genomic composition	Accession No.	Provenance	Provider
Th. elongatumTh. elongatumTh. elongatumTh. elongatumTh. bessarabicumTh. bessarabicum	2n = 142n = 142n = 142n = 142n = 142n = 14	E^eE^eE^eE^eE^eE^eE^eE^eE^bE^b ( = JJ)E^bE^b ( = JJ)	PI 531719PI 531717PI 531718PI 153179PI 531711W6 21890	FranceFranceTunisiaTurkeyEstoniaUkraine	[a][a][a][a][a][a]
Th. intermediumTh. intermediumTh. intermediumTh. intermediumTh. intermediumTh. intermediumTh. intermediumTh. intermediumTh. intermediumTh. intermedium	2n = 422n = 422n = 422n = 422n = 422n = 422n = 422n = 422n = 422n = 42	E^eE^eE^bE^bStStE^eE^eE^bE^bStStE^eE^eE^bE^bStStE^eE^eE^bE^bStStE^eE^eE^bE^bStStE^eE^eE^bE^bStStE^eE^eE^bE^bStStE^eE^eE^bE^bStStE^eE^eE^bE^bStStE^eE^eE^bE^bStSt	PI 401225PI 401178PI 401183PI 401196PI 383575PI 547332PI 547335PI 547316PI 547315PI 547319	IranIranIranIranTurkeyTurkeyPolandRussian FederationRussian FederationRussian Federation	[a][a][a][a][a][a][a][a][a][a]
Octoploid trititrigia	2n = 562n = 562n = 562n = 542n = 562n = 562n = 562n = 562n = 562n = 56	AABBDDEE40W + 8J+ 8J^sAABBDDEEAABBDD+ 2St+ 4J+ 4J^s + 2St-J^s40W + 4St+ 2Js+ 2Js-W + 2S-Js+ 2St-Js+ 2S-J+ 2W-Js40W + 8St+ 4J^s + 2J+ 2J-St40W + 6St+ 8EAABBDD+ 8St+ 4E+ 2St-E40W + 2St+ 12E+ 2St-EAABBDD+ 12J^s + 2J	KD20041XY6937430Zhong2Zhong5TE70447045704778784	USAChinaChinaChinaChinaChinaChinaChinaChinaChina	[b][c][c][d][d][e][f][f][f][g]
Th. elongatumTh. elongatumTh. elongatumTh. elongatumTh. elongatumTh. elongatumTh. elongatumTh. elongatumTh. elongatumTh. elongatum	2n = 702n = 702n = 702n = 702n = 702n = 702n = 702n = 702n = 702n = 70	- (!)- (!)- (!)- (!)- (!)- (!)- (!)- (!)- (!)- (!)	PI 315352PI 308592PI 297871PI 283164PI 276399PI 368851PI 251443PI 401008PI 401009PI 401010	Former Soviet UnionItalyArgentinaChinaGermanyUSATurkeyTurkeyTurkeyTurkey	[a][a][a][a][a][a][a][a][a][a]
Wheat-Th. elongatum/Th. intermedium additions	2n = 442n = 442n = 442n = 442n = 442n = 442n = 442n = 442n = 442n = 442n = 442n = 442n = 44	AABBDD+ one pair 1EAABBDD+ one pair 2EAABBDD+ one pair 3EAABBDD+ one pair 4EAABBDD+ one pair 5EAABBDD+ one pair 6EAABBDD+ one pair 7EAABBDD+ one pair S-J^sAABBDD+ one pair S-J^sAABBDD+ one pair SAABBDD+ one pair W-J^sAABBDD+ one pair W-J^sAABBDD+ one pair S-J^s	KD20061KD20062KD20063KD20064KD20065KD20066KD20067Z1Z2Z3Z4Z5Z6	USAUSAUSAUSAUSAUSAUSAChinaChinaChinaChinaChinaChina	[b][b][b][b][b][b][b][a][a][a][a][a][a]
Wheat-Thinopyrum intermedium ssp. trichophorum introgressions	2n = 562n = 56—	40W + 9St+ 4J^s + 3J40W + 8St+ 6J^s + 2J—	1908Q156Q157-Q227	ChinaChinaChina	[e1][e1][e]
Ps. SpicataS. cerealeS. africanumS. silvestreD. breviaristatumD. villosumH. vulgareAe.comosaAe.umbellulataAe.bicornisAe. cylindricaAg.cristatumA.retrofractumEremopyrumAe. tauschiiT. monococcumT. turgidumT. timopheeviiT .teresT. aestivumT. aestivumT. aestivumT. aestivum	2n = 142n = 142n = 142n = 142n = 282n = 142n = 142n = 142n = 142n = 142n = 282n = 282n = 142n = 142n = 142n = 142n = 282n = 282n = 282n = 422n = 422n = 422n = 42	StStRRR^aR^aR^sR^sV^bV^bV^bV^bVVHHMMUUS^bS^bCCDDPPPPWWFFDDAAAABBAAGGA^uA^uDDAABBDDAABBDD (1RS/1BL)AABBDD (1RS/1BL)AABBDD (1RS/1BL)	PI 232131SCND1377CN41211CN41954KD19791KD19801CN41966PI 317400PI 227339CIae 70PI 374320PI 578520PI 547363PI 227343KD20042PI 427328PI 308878PI 297326KD1988CSCN12CN18CN17	USAChinaSouth AfricaAustriaMoroccoMoroccoChinaAfghanistanIranUnknownYugoslaviaCanadaAustraliaIranChinaIraqSpainFormer Soviet UnionChinaChinaChinaChinaChina	[a][e][h][h][a][i][i][a][a][a][a][a][a][a][e][a][a][a][j][h][i][i][i]

Tab.1 Materials used in this study

Tab.2 Information about the primers designed in this study

Fig.1 DNA amplification patterns of , , Hexaploid Trititrigia and controls with primers D14F and D14R. M: Marker(DL1000); Lanes 1 to 10 are PI 531719, PI 531717, PI 531711, W6 21890, PI 401178, PI 401225, PI 383575, PI 547332, PI 547335 and PI 547316, respectively; Lanes 11 to 17 are Chinese Spring, , subsp, , and wheat varieties CN18, CN12, respectively; Lane 18 is blank control. Arrow showed the target PCR bands.

Fig.2 DNA amplification patterns of , Octoploid and controls with primers D14F and D14R. M: Marker(DL4500); Lanes 1 to 12 are , Chinese spring- amphiploid, Chinese spring- amphiploid, Wheat- ssp amphiploid and Octoploid 7044, 7045, 7047, 78784, 7430, Zhong2, Zhong5 and XY693, respectively; Lanes 13 to 17 are CS, CN12, CN18 and CN17. Lane 18 is blank control. Arrow showed the target PCR bands.

Fig.3 DNA amplification patterns of wheat- additions and controls with primers D14F and D14R. M: Marker(DL4500); Lanes 1 to 7 are Chinese spring- additions CSDA1E-CSDA7E; Lanes 8 to 13 are wheat- additions Z1-Z6; Lanes 14 to 17 are CS, CN12, CN18 and CN17, respectively; Lane 18 is blank control. Arrow showed the target PCR bands.

Fig.4 Fluorescence in situ hybridization analysis using 4I (A) and genomic DNA (B-D) as probe and without blocking DNA used on mitotic metaphase chromosomes of Chinese spring- amphiploid (A) and wheat- ssp introgressions 1908 (B), Q156 (C) and Q168 (D). ? in Fig. 4D means unknown chromosome due to chromosome overlap or weak hybridization signal.

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