Morphological characteristics and molecular identification of <em>Kappaphycus alvarezii</em>

ZHAO Su-fen; HE Pei-min

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ZHAO Su-fen, HE Pei-min. Morphological characteristics and molecular identification of Kappaphycus alvarezii[J]. Haiyang Xuebao, 2010, 32(6): 157-166.

Citation:

ZHAO Su-fen, HE Pei-min. Morphological characteristics and molecular identification of Kappaphycus alvarezii[J]. Haiyang Xuebao, 2010, 32(6): 157-166.

ZHAO Su-fen, HE Pei-min. Morphological characteristics and molecular identification of Kappaphycus alvarezii[J]. Haiyang Xuebao, 2010, 32(6): 157-166.

Citation:

ZHAO Su-fen, HE Pei-min. Morphological characteristics and molecular identification of Kappaphycus alvarezii[J]. Haiyang Xuebao, 2010, 32(6): 157-166.

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Morphological characteristics and molecular identification of Kappaphycus alvarezii

ZHAO Su-fen¹,
HE Pei-min^2
,

1.
College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China; Fisheries College, Guangdong Ocean University, Zhanjiang 524025, China
2.
Fisheries College, Guangdong Ocean University, Zhanjiang 524025, China

Received Date: 2009-08-27
Rev Recd Date: 2009-11-10

Abstract

Abstract

Three lines (Line XC-03 of Kappaphycus alvarezii from Lingshui coast, Line LD-02 and Line LD-06 from Sanya coast) of eucheumatoids algae with different shapes cultivated in Hainan Province, China were identified by morphological characteristics and molecular phylogenetic analysis, and were compared with Line TSNY-05 of Eucheuma denticulatum from Tanzania. There were differences in the branch shapes, length and distributing and tuber numbers, the cell sizes of epidermis, cortex and medulla part, the counts of cortex and the thickness of the colloid among three lines (Line XC-03, Line LD-02 and Line LD-06). But molecular identification indicated that Line XC-03, Line LD-02 and Line LD-06 were same species, K. alvarezii. The genome DNA was extracted from four lines by CTAB method. 5 polymorphic primers were screened out of 30 primers for random amplified polymorphic DNA (RAPD) analyses and 39 primers for inter-simple sequence repeat (ISSR) analyses, respectively. 12 individuals were used for molecular analysis respectively, and 39 loci (RAPD) and 49 loci (ISSR) were obtained. The fragments for RAPD and ISSR markers were from 300 to 3 000 bp or from 400 to 3 000 bp, respectively. The PCR amplified products of the internal transcribed spacer (ITS) were sequenced by the ABI PRISM 3730 genetic analyzer. The sequence data were blasted and aligned, and phylogeny trees were constructed by neighbour-joining and UPGMA analyses using MEGA 4.0. The results showed that the similarity of the ITS sequence was 99.69%~99.85% among Line XC-03, Line LD-02 and Line LD-06. There was one base transition and one base transversion between Line XC-03 and Line LD-02, and there was only one base transition between Line XC-03 and Line LD-06 and one base transversion between Line LD-02 and Line LD-06. While the similarity of the ITS sequence was 41.28% between K. alvarezii.and E. denticulatum. And RAPD, ISSR and ITS analysis further indicated that the pairwise genetic distance between Line XC-03, Line LD-02 and Line LD-06 was from 0.122 9 to 0.153 5 (RAPD), from 0.068 8 to 0.112 4(ISSR) and from 0.002 to 0.005 (ITS), respectively, with software Popgen 32 and MEGA 4.0. With Neighbor-Joining and UPGMA analyses, it indicated that the relationship between Line XC-03 and LD-06 was closer than that between Line XC-03 and Line LD-02. However, the pairwise genetic distance was above 0.9 between E. denticulatum and K. alvarezii, and it showed that the relationship was very far between E. denticulatum.and K. alvarezii. These results provide a basis for the species identification and genetics breeding in K. alvarezii cultivation.
- Kapphphycus alvarezii,
- morphological characteristics,
- RAPD,
- ISSR,
- ITS

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References(37)

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