Yao, Fang-Jie
(College of Horticulture, Jilin Agricultural University)
,
Lu, Li-Xin
(Engineering Research Center of Chinese Ministry of Education for Edible and Medicinal Fungi, Jilin Agricultural University)
,
Wang, Peng
(Engineering Research Center of Chinese Ministry of Education for Edible and Medicinal Fungi, Jilin Agricultural University)
,
Fang, Ming
(College of Horticulture, Jilin Agricultural University)
,
Zhang, You-Min
(College of Horticulture, Jilin Agricultural University)
,
Chen, Ying
(Engineering Research Center of Chinese Ministry of Education for Edible and Medicinal Fungi, Jilin Agricultural University)
,
Zhang, Wei-Tong
(Engineering Research Center of Chinese Ministry of Education for Edible and Medicinal Fungi, Jilin Agricultural University)
,
Kong, Xiang-Hui
(Engineering Research Center of Chinese Ministry of Education for Edible and Medicinal Fungi, Jilin Agricultural University)
,
Lu, Jia
(College of Horticulture, Jilin Agricultural University)
,
Honda, Yoichi
(Graduate School of Agriculture, Kyoto University)
The fruiting body pattern is an important agronomic trait of the edible fungus Auricularia auricula-judae, and an important breeding target. There are two types of fruiting body pattern: the cluster type and the chrysanthemum type. We identified the fruiting body pattern of 26 test strains, and then...
The fruiting body pattern is an important agronomic trait of the edible fungus Auricularia auricula-judae, and an important breeding target. There are two types of fruiting body pattern: the cluster type and the chrysanthemum type. We identified the fruiting body pattern of 26 test strains, and then constructed two different near-isogenic pools. Then, we developed sequence characterized amplified region (SCAR) molecular markers associated with the fruiting body pattern based on sequence-related amplified polymorphism (SRAP) markers. Ten different bands (189-522 bp) were amplified using 153 pairs of SRAP primers. The SCAR marker "SCL-18" consisted of a single 522-bp band amplified from the cluster-type strains, but not the chrysanthemum strains. This SCAR marker was closely associated with the cluster-type fruiting body trait of A. auricula-judae. These results lay the foundation for further research to locate and clone genes controlling the fruiting body pattern of A. auricula-judae.
The fruiting body pattern is an important agronomic trait of the edible fungus Auricularia auricula-judae, and an important breeding target. There are two types of fruiting body pattern: the cluster type and the chrysanthemum type. We identified the fruiting body pattern of 26 test strains, and then constructed two different near-isogenic pools. Then, we developed sequence characterized amplified region (SCAR) molecular markers associated with the fruiting body pattern based on sequence-related amplified polymorphism (SRAP) markers. Ten different bands (189-522 bp) were amplified using 153 pairs of SRAP primers. The SCAR marker "SCL-18" consisted of a single 522-bp band amplified from the cluster-type strains, but not the chrysanthemum strains. This SCAR marker was closely associated with the cluster-type fruiting body trait of A. auricula-judae. These results lay the foundation for further research to locate and clone genes controlling the fruiting body pattern of A. auricula-judae.
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제안 방법
The SRAP markers are highly stable and polymorphic, but the conversion rate is low [30]. In this study, SRAP markers were used to screen the polymorphisms related to fruiting body traits. From the successful SRAP markers, we generate a stable 522-bp SCAR marker designated as SCL-18.
In this study, specific polymorphic fragments were amplified using many SRAP marker primers, and then transformed into a single, stable SCAR marker, which differentiated between CL and CH strains at the DNA level. This provides an effective method for molecular marker-assisted breeding for the fruiting body pattern in A.
In this study, the SCAR marker SCL-18 related to the CL type was successfully screened from hundreds of pairs of SRAP primers. Compared with protein bands or isozyme markers, DNA markers are more conservative and less susceptible to environmental effects [29].
Specific bands associated with the fruiting body pattern were screened, excised, and purified with an AXYGEN DNA Gel Extraction Kit (Axygen, Union City, CA). The extracted DNA was ligated into the pEASYV®-T5 zero cloning vector (TransGen Biotech Co.
The extracted DNA was ligated into the pEASYV®-T5 zero cloning vector (TransGen Biotech Co., Beijing, China), and the positive clones were screened for DNA sequencing.
The sequencing results were used to design fruiting body pattern-specific SCAR primers to identify fruiting body patterns in the verified strains shown in Table 1. The primers were designed with Primer 3 software [25].
B1–B20 are nationally accredited varieties commonly cultivated in China. These verified strains were used to test the SCAR markers developed in this study. The fruiting body patterns of all strains were confirmed by a fruiting test [4].
대상 데이터
Data analysis and sequence alignments were conducted using DNAMANV® version 5.2.9 (Lynnon Bio Soft, San Ramon, CA) and the GenBank database(https://www.ncbi.nlm.nih.gov/).
In total, 153 pairs of SRAP primers were used to amplify fragments from the CL-type and CH-type isogenic lines. Ten pairs of primers could distinguish the fruiting body pattern of all the monokaryotic strains.
이론/모형
Total genomic DNA was extracted from F1 monokaryotic strains by a modified CTAB method [21,22]. The concentration of DNA from each strain was diluted to 50 mg/lL.
후속연구
The sequences of the specific fragments obtained in this study were used as BLASTN queries to search the GenBank database, but there were no homologous sequences in the database. In future research, we will try to locate the trait loci on the SSR genetic linkage map of A. auricula-judae. This will allow us to explore genetic regulation mechanisms and genes of interest through the linkage relationships with other traits and/or molecular markers.
1 Yao FJ , Bian YB. Graphic illustration on key cultivation techniques of Auricularia auricular-judae . Beijing : China Agricultural Press ; 2011 .
2 Yao FJ , Zhang YM , Lu LX , et al Research progress on genetics and breeding of Auricularia auricula-judae . J Fungal Res . 2015 ; 13 : 125 – 128 .
3 Chen Y , Yao FJ , Zhang YM , et al Numerical classification of cultivated germplasm of Auricularia auricular-judae . Mycosystema . 2014 ; 33 : 984 – 996 .
4 Chen Y , Yao FJ , Zhang YM , Fang M. Study on esterase isozyme of cultivated germplasm on Auricularia auricula-judae (Bull.) Quél In: The 10th National Academic Symposium on Edible Fungi; 2014 Mar 21–23 ; Beijing, China. Beijing : Mycological society of China ; 2014 p. 75 – 80 .
5 Wang P , Yao FJ. Studies on germplasm resources evaluation by SSR on Auricularia auricular-judae In: The 11th Academic Symposium on Medicinal Fungi; 2016 Oct 27–30 ; Chengdu, China. Beijing : China edible fungi association ; 2016 .
6 Chen Y. Studies on numerical classification of germplasm, directional breeding model construction and differential gene expression analysis of degeneration strain by cDNA-AFLP on Auricularia auricular-judae [dissertation] . Changchun : Jilin Agricultural University ; 2014 .
7 Chen Y , Yao FJ , Zhang YM , et al A new Auricularia auricula cultivar ‘Jihei 3’ . Acta Hortic Sinica . 2014 ; 41 : 1751 – 1752 .
8 Fang M , Yao FJ , Wang XE , et al A new Auricularia auricula cultivar ‘Jihei 2’ . Acta Hortic Sinica . 2013 ; 40 : 1215 – 1216 .
9 Bhagyawant SS. RAPD-SCAR markers: an interface tool for authentication of traits . J Biosci Med . 2016 ; 4 : 1 – 9 .
10 Cunha CMS , Hinz RH , Pereira A , et al A SCAR marker for identifying susceptibility to Fusarium oxysporum f. sp. Cunense in banana . Sci Hortic . 2015 ; 191 : 108 – 112 .
11 Khan M , Pan Y , Iqbal J. Development of an RAPD-based SCAR marker for smut disease resistance in commercial sugarcane cultivars of Pakistan . Crop Prot . 2017 ; 94 : 166 – 172 .
12 Loftus M , Bouchti KL , Robles C , et al Use of SCAR marker for cap color in Agaricus bisporus breeding programs . Mush Sci . 2000 ; 15 : 201 – 205 .
13 Kong WS , Kim GH , Kim KH , et al Molecular marker related to fruitbody color of Flammulina velutipes . Mycobiology . 2004 ; 32 : 6 – 10 .
14 Lee CY , Park JE , Lee J , et al Development of new strains and related SCAR markers for an edible mushroom, Hypsizygus marmoreus . FEMS Microbiol Lett . 2012 ; 327 : 54 – 59 . 22093038
15 Li DQ , Wang J. Screening of molecular markers associated with degeneration of Volvariella volvacea strains . J Northwest A&F Univ (Nat. Sci. Ed) . 2015 ; 43 : 195 – 201 .
16 Kim SY , Kim KH , Im CH , et al Identification of degenerate nuclei and development of a SCAR marker for Flammulina velutipes . PLoS One . 2014 ; 9 : e107207 . 25221949
18 Li D , Liu Y , Wang P , et al Development of SCAR markers to determine the mating types of Lepista nuda protoplast monokaryons . Curr Microbiol . 2014 ; 68 : 536 – 542 . 24352297
19 Zhao Y , Lin F , Yan SY. Establishment and application of an efficient cross breeding method assisted by molecular markers of Volvariella volvacea . Microbiology China . 2015 ; 42 : 1165 – 1174 .
20 Liu GJ. Study on incompatible factor and breeding of Auricularia auricular [dissertation] . Changchun : Jilin Agricultural University ; 2011 .
21 Murray MG , Thompson WF. Rapid isolation of high molecular weight plant DNA . Nucleic Acids Res . 1980 ; 8 : 4321 – 4326 . 7433111
22 Wang XE. Analysis of Genome and lignocellulose degradation genes and new cultivar breeding of Auricularia auricular [dissertation] . Changchun : Jilin Agricultural University ; 2013 .
23 Li G , Quiros CF. Sequence-related amplified polymorphism (SRAP), a new marker system based on a simple PCR reaction: its application to mapping and gene tagging in Brassica . Theor Appl Genet . 2001 ; 103 : 455 – 461 .
24 Lin ZX , Zhang XL , Nie YC , et al Construction SRAP genetic linkage map in cotton . Chin Sci Bullet . 2003 ; 15 : 1676 – 1679 .
25 Rozen S , Skaletsky H Primer3 on the WWW for general users and for biologist programmers . Methods Mol Biol . 2000 ; 132 : 365 – 386 . 10547847
26 Baars JJP , Sonnenberg ASM , Mikosch TSP , et al Development of a sporeless strain of oyster mushroom Pleurotus ostreatus . Indian J Pharm Sci . 2000 ; 74 : 588 – 591 .
27 Chen Y. Study on the diversity of cultured Auricularia auricular germplasm resources and the establishment of core collection [dissertation] . Changchun : Jilin Agricultural University ; 2010 .
28 Wan JN. Study on effect and mechanism of small hole-fruiting method on quality of Auricularia auricular [dissertation] . Changchun : Jilin Agricultural University ; 2009 .
29 Chang ST , Lin FC. Genetics and breedings of edible mushroom . Beijing : China Agriculture Press ; 1997 .
30 Callac P , Moquet F , Imbernon M , et al Evidence for PPC1 a determinant of the pilei-pellis color of Agaricus bisporus fruiting bodies . Fungal Genet Biol . 1998 ; 23 : 181 – 188 . 9578631
31 Moquet F , Desmerger C , Mamoun M , et al A quantitative trait locus of Agaricus bisporus resistance to Pseudomonas tolaasii is closely linked to natural cap color . Fungal Genet Biol . 1999 ; 28 : 34 – 42 . 10512670
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