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2024 Vol.42, Issue 6

Research Article

Development of SSRs to Identify Self-fertilized and Nucellar Seedlings in Common Trifoliate Orange and Flying Dragon

일반 탱자 및 ‘비룡’ 탱자에서 자가수정 및 주심배 유래의 구분 SSR 선발

Sung Yeon Kim1
,
Eun Ui Oh1
,
Ho Bang Kim2
,
Kwan Jeong Song1*
김 성연1
,
오 은의1
,
김 호방2
,
송 관정1*
1Department of Horticultural Science, Graduate School, SARI, Jeju National University, Jeju 63243, Korea
2Department of Environmental Horticulture, School of Equine Science and Horticulture, Cheju Halla University, Jeju 63092, Korea
1제주대학교 대학원 원예학과
2한라대학교 생명자원학부 환경원예과
*Corresponding Author
31 December 2024. pp. 749-758
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Abstract

Most citrus rootstocks in Korea are seedlings of trifoliate orange. They are mainly known to have a nucellar origin, but at a low rate, those with self-fertilized origins are mixed in, and it is difficult to distinguish between the two types visually. If some self-fertilized seedlings are thus mixed, nonuniformity of citrus planting stocks would appear. Therefore, this study was conducted to evaluate the frequency of self-fertilized seedlings and the uniformity of seedlings using with DNA markers that identify nucellar and self-fertilized seedlings in trifoliate oranges. Twenty SSRs were selected and determined to be heterozygous in the twisted ‘Flying Dragon’ (Poncirus trifoliata cv. Flying Dragon) and homozygous in the straight ‘Flying Dragon’, originating from self-fertilization. A total of seven representative SSRs, capable of double checking in a single plant, were finally selected. As a result of an origin analysis by applying the selected SSRs to a common population of trifoliate oranges, more of those with self-fertilized origins were detected from small plants than large plants when two seedlings were grown from one seed. In particular, the largest plant did not have a self-fertilized origin when several plants were grown from one seed sown in vitro. Also, when the frequency of self-fertilized seedlings was evaluated during the first year and second year in a nursery, a self-fertilized origin was detected at a rate of 4.0% in large seedlings and 26% in small seedlings in the first year after sowing. Plants with poor growth were rogued and degenerated in the second year after sowing, and their self-fertilized origins were detected at a rate of 5.5%. The results here suggests that the selected SSR markers would be helpful to improve the uniformity of the production of citrus planting stocks and would enable the detection of nonuniform growth in citrus plants.

Keywords
citrus rootstocks
DNA marker
nucellar embryo
polyembryony
Poncirus trifoliata

국내 감귤 대목으로는 거의 대부분 탱자 실생묘를 사용하고 있다. 탱자 실생묘는 대부분 주심배 유래이고, 자가수정 유래가 낮은 비율로 혼재되어 있으나 육안 구별은 거의 불가능하다. 자가수정 유래의 탱자묘가 일부 혼재하게 되면 감귤 묘목의 불균일성이 나타나게 된다. 그러므로 탱자 실생묘에서 자가수정 및 주심배 유래를 구분하는 SSR 선발을 통해 자가수정 유래의 빈도를 평가하고 묘목의 균일성을 높이는데 활용하고자 본 연구를 수행하였다. 뒤틀림 외형에서 이형접합체로, 그리고 자가수정 유래의 비뒤틀림 외형에서는 동형접합체로 나타내는 SSR 20개를 선발하였으며, 이 중 한 개체에서 2개이상 확인 가능한 7개를 최종 선발하였다. 선발된 SSR을 일반 탱자에 적용하여 유래 분석을 실시한 결과, 한 종자에서 두 개의 묘가 발생했을 때 큰 식물체보다 작은 식물체에서 더 많은 자가수정 유래가 확인됐으며, 기내파종되어 여러 개체를 발생시켰을 때 가장 큰 개체에서는 자가수정이 확인되지 않았다. 한편 육묘장의 파종 1년차 실생 집단 중 키가 큰 실생묘에서는 4.0%, 작은 실생묘에서는 26.0%로 자가수정 유래가 확인되었다. 생육이 불량한 개체를 분리 도태시킨 파종 2년차 정식 집단에서는 자가수정이 6.6%의 비율로 나타났다. 본 연구를 통해 선발된 SSR을 활용하면 감귤 묘목 생산의 균일성을 높임은 물론 감귤에서 생육이 불균일한 개체에서 자가수정 유래 탱자 대목의 영향 여부에 대해 확인이 가능하게 될 것이다.

키워드
감귤 대목
DNA 마커
주심배
다배성
Poncirus trifoliata
References
1

Aleza P, Juárez J, Ollitrault P, Navarro L (2010) Polyembryony in non-apomictic citrus genotypes. Ann Bot 106:533-545. doi:10.1093/aob/mcq148

10.1093/aob/mcq14820675656PMC2944972
2

Anderson CM, Castle WS, Moore GA (1991) Isozymic identification of zygotic seedlings in swingle citrumelo Citrus paradisi X Poncirus trifoliata nursery and field populations. J Amer Soc Hort Sci 116:322-326. doi:10.21273/JASHS.116.2.322

10.21273/JASHS.116.2.322
3

Andrade-Rodriguez M, Montecillo C, Villegas-Monter A, Gutierrez-Espinosa MA, Carrillo-Castaneda G, Garcia-Velazquez A (2005) Polyembryony and RAPD markers for identification of zygotic and nucellar seedlings in Citrus. Agrociencia 39:371-383. doi:10.1590/S0100-204X2004000600006

10.1590/S0100-204X2004000600006
4

Chae CW, Yun SH, Park JH, Kim MJ, Han SG, Kang SB, Koh SW, Han SH (2013) Improvement of seed germination in a spontaneous autotetraploid of poncirus and chlorophyll fluorescence of seedlings in salt stress. J Life Sci 23:1079-1087. doi:10.5352/JLS.2013.23.9.1079

10.5352/JLS.2013.23.9.1079
5

Cheng FS, Roose ML (1995) Origin and inheritance of dwarfing by the citrus rootstock Poncirus trifoliata 'Flying Dragon'. J Amer Soc Hort Sci 120:286-291. doi:10.21273/JASHS.120.2.286

10.21273/JASHS.120.2.286
6

Cheng YJ, Guo WW, Yi HL, Pang XM, Deng X (2003) An efficient protocol for genomic DNA extraction from Citrus species. Plant Mol Biol Rep 21:177-178. doi:10.1007/BF02774246

10.1007/BF02774246
7

Doyle JJ, Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus 12:13-15. doi:10.2307/2419362

10.2307/2419362
8

Fukui K (2020) Genetic and breeding the second edition of the plant. Worldscience, Seoul, Korea

9

Gill MIS, Chawla N, Chopra HR (2002) Isozyme characterization and indexing of in vitro raised nucellars in Citrus rootstocks. Indian J Hortic 59:122-126

10

Kamiri M, Stiff M, Costantino G, Dambier D, Kabbage T, Ollitrault P, Froelicher Y (2018) Preferential homologous chromosome pairing in a tetraploid intergeneric somatic hybrid (Citrus reticulata + Poncirus trifoliata) revealed by molecular marker inheritance. Front Plant Sci 9:1557. doi:10.3389/fpls.2018.01557

10.3389/fpls.2018.0155730450106PMC6224360
11

Kaul H, Sidhu GS, Sarao NK, Singh R, Singh G (2022) Assessment of genetic diversity of mandarin cultivars grown in major citrus regions of world using morphological and microsatellite markers. Hortic Environ Biotechnol 63:25-437. doi:10.1007/s13580-021-00404-4

10.1007/s13580-021-00404-4
12

Kepiro JL, Roose ML (2007) Nucellar embryony. Citrus Genetics, Breeding and Biotechnology. CABI, Wallingford, UK. doi:10.1079/9780851990194.0141

10.1079/9780851990194.0141
13

Khan IA, Roose ML (1988) Frequency and characteristics of nucellar and zygotic seedlings in three cultivars of trifoliate orange. J Amer Soc Hort Sci 113:105-110. doi:10.21273/JASHS.113.1.105

10.21273/JASHS.113.1.105
14

Kim JH, Sato M, Wakana A, Takamatsu F, Skai K, Shigyo M, Masuda JI (2021a) Genotyping of Citrus accessions with S9 and/or S10 alleles for self-incompatibility and their allelic distribution. Hortic Sci Technol 39:807-822. doi:10.7235/HORT.20210071

10.7235/HORT.20210071
15

Kim MS, Kim SH, Kim HB, Park YC, Song KJ (2020) Some factors affecting the efficiency of hybrid embryo rescue in the 'Shiranuhi' mandarin. Hortic Sci Technol 38:271-281. doi:10.7235/HORT.20200026

10.7235/HORT.20200026
16

Kim SR, Jo NS, Gil JS, Koo SC, Um YR, Hong CP, Park SG, Kim OT, Kim SC, et al. (2021b) Development of genome-wide simple sequence repeat markers in Codonopsis lanceolata using next-generation sequencing. Hortic Environ Biotechnol 62:985-993. doi:10.1007/s13580-021-00389-0

10.1007/s13580-021-00389-0
17

Kim SS, Kim HB, Park KJ, Hyun JW, Choi CW, Joa JH, Jin SB, Kim ES, Han SG (2021c) Development of plastid InDel markers to discriminate lemons from other citrus groups. Hortic Sci Technol 39:637-644. doi:10.7235/HORT.20210057

10.7235/HORT.20210057
18

Kishore K, Monika N, Rinchen D, Lepcha B, Pandey B (2012) Polyembryony and seedling emergence traits in apomictic Citrus. Sci Hortic 138:101-107. doi:10.1016/j.scienta.2012.01.035

10.1016/j.scienta.2012.01.035
19

Koh HJ, Kwon SY, Thomson M (2015) Current technologies in plant molecular breeding: A Guide Book of Plant Molecular Breeding for Researchers. Springer, New York, USA. doi:10.1007/978-94-017-9996-6

10.1007/978-94-017-9996-6
20

Koltunow AM (1993) Apomixis: Embryo sacs and embryos formed without meiosis or fertilization in ovules. Plant Cell 5:1425-1437. doi:10.2307/3869793

10.2307/386979312271038PMC160373
21

Korean Statistical Information Service (KOSIS) (2021) https://kosis.kr. Accessed 2 December 2022

22

Mademba-Sy F, Lemerre-Desprez Z, Lebegin S (2012) Use of Flying Dragon trifoliate orange as dwarfing rootstock for citrus under tropical climatic conditions. HortScience 47:11-17. doi:10.21273/HORTSCI.47.1.11

10.21273/HORTSCI.47.1.11
23

Martínez-Cuenca MR, Primo-Capella A, Forner-Giner MA (2021) Screening of 'King' mandarin (Citrus nobilis Lour) × Poncirus trifoliata ((L.) Raf.) hybrids as salt stress-tolerant citrus rootstocks. Hortic Environ Biotechnol 62:337-351. doi:10.1007/s13580-020-00291-1

10.1007/s13580-020-00291-1
24

Ministry of Agriculture, Food and Rural Affairs (MAFRA) (2021) https://kass.mafra.go.kr. Accessed 2 December 2022

25

Moore GA, Castle WS (1988) Morphological and isozymic analysis of open-pollinated citrus rootstock populations. J Hered 79:59-63. doi:10.1093/oxfordjournals.jhered.a110448

10.1093/oxfordjournals.jhered.a110448
26

Roose ML, Prechtl SN (1988) Identification and performance of citrus trees on nucellar and zygotic rootstocks. J Amer Soc Hort Sci 113:100-105. doi:10.21273/JASHS.113.1.100

10.21273/JASHS.113.1.100
27

Villegas-Monter A, Matinez-Ochoa EDC, Andrade-Rodriguez M, Villegas-Velazquez I (2022) Citrus polyembryony. In Advances in Citrus Production and Research. London, UK: IntechOpen. doi:10.5772/intechopen.105994

10.5772/intechopen.105994
28

Woo JK, Yun SH, Yi KU, Park YC, Lee HY, Kim MJ, Lee Y, Song KJ, Kim HB (2020) Identification of citrus varieties bred in korea using microsatellite markers. Hortic Sci Technol 38:374-384. doi:10.7235/HORT.20200036

10.7235/HORT.20200036
29

Yun JU, Yang HB, Jung YH, Yun SH, Kim KS, Kim CS, Song KJ (2007) Identification of zygotic and nucellar mandarin seedlings using randomly amplified polymorphic DNA. Hortic Environ Biotechnol 48:171-175

Information
  • Publisher :KOREAN SOCIETY FOR HORTICULTURAL SCIENCE
  • Publisher(Ko) :한국원예학회
  • Journal Title :Horticultural Science and Technology
  • Journal Title(Ko) :원예과학기술지
  • Volume : 42
  • No :6
  • Pages :749-758
  • Received Date : 2023-11-06
  • Revised Date : 2023-12-13
  • Accepted Date : 2024-01-08
  • DOI :https://doi.org/10.7235/HORT.20240051
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