Method for identifying cytoplasm type of three-line sterile rice by detecting chloroplast DNA (deoxyribose nucleic acid)

Abstract

The invention discloses a method for identifying the cytoplasm type of three-line sterile rice by detecting chloroplast DNA (deoxyribose nucleic acid). The method comprises the steps of: taking rice leaves as material, extracting chloroplast DNA in the leaves, carrying out PCR (polymerse chin rection) amplification by using chloroplast primers cp1, cp2, cp3 and cp9, cloning and sequencing the amplified product, and comparing the result with chloroplast DNA molecular fingerprinting of the three-line sterile rice so as to simply and efficiently identify the cytoplasm type of the three-line sterile rice.

Description

technical field

[0001] The invention relates to a method for detecting chloroplast DNA to identify the cytoplasm type of three-line sterile rice. Background technique

[0002] In 1966, Japanese scholar Shinjo Changtomo bred a BT-type male sterile line (selected with Indian indica rice variety Chinsuran Boro II as cytoplasmic donor and japonica rice variety as nuclear donor) controlled by both cytoplasm and nucleus. In 1970, Yuan Longping, Li Bihu and others discovered a male sterile plant in Hainan wild rice, and bred it into a "wild defeat" type three-line sterile line through nuclear replacement. Subsequently, the Honglian type three-line male sterile line was bred by crossing Hainan Hongmans wild rice and indica variety Liantangzao. Later, the K-type, Gang-type, Yinshui-type and so on were bred. Since these cytoplasms have no cytoplasmic marker traits except those related to male sterility, it is impossible to distinguish their cytoplasms by common biological traits. ...

Examples

Embodiment Construction

[0010] In the present invention, 32 typical indica rice and typical japonica rice cytoplasms were used as controls, and the chloroplast DNA sequences of 17 three-line sterile lines from different sources were analyzed, and it was found that different types of sterile cytoplasms had characteristic bases or base sequences, and the chloroplast primers The amplified sequence difference of cp1, cp2, cp3 and cp9 established three chloroplast DNA molecular fingerprints of wild type, red lotus type and BT type.

[0011] Using conventional rice Nipponbare as a control, PCR amplification products of chloroplast DNA of three-line male sterile lines such as V20A were cloned and sequenced, and the sequence results are shown in Table 1.

[0012] Table 1

[0013]

[0014] Note: "-" means missing the sequence.

[0015] Chloroplast primers described in Table 1

[0016] cp1: forward sequence 5-gtggacctgactccttgaa-3, reverse sequence 5-agccgaggtcgtggtaa-3;

[0017] cp2: forward sequence 5-...

technical field

[0001] The invention relates to a method for detecting chloroplast DNA to identify the cytoplasm type of three-line sterile rice. Background technique

[0002] In 1966, Japanese scholar Shinjo Changtomo bred a BT-type male sterile line (selected with Indian indica rice variety Chinsuran Boro II as cytoplasmic donor and japonica rice variety as nuclear donor) controlled by both cytoplasm and nucleus. In 1970, Yuan Longping, Li Bihu and others discovered a male sterile plant in Hainan wild rice, and bred it into a "wild defeat" type three-line sterile line through nuclear replacement. Subsequently, the Honglian type three-line male sterile line was bred by crossing Hainan Hongmans wild rice and indica variety Liantangzao. Later, the K-type, Gang-type, Yinshui-type and so on were bred. Since these cytoplasms have no cytoplasmic marker traits except those related to male sterility, it is impossible to distinguish their cytoplasms by common biological traits. ...

Examples

Embodiment Construction

[0010] In the present invention, 32 typical indica rice and typical japonica rice cytoplasms were used as controls, and the chloroplast DNA sequences of 17 three-line sterile lines from different sources were analyzed, and it was found that different types of sterile cytoplasms had characteristic bases or base sequences, and the chloroplast primers The amplified sequence difference of cp1, cp2, cp3 and cp9 established three chloroplast DNA molecular fingerprints of wild type, red lotus type and BT type.

[0011] Using conventional rice Nipponbare as a control, PCR amplification products of chloroplast DNA of three-line male sterile lines such as V20A were cloned and sequenced, and the sequence results are shown in Table 1.

[0012] Table 1

[0013]

[0014] Note: "-" means missing the sequence.

[0015] Chloroplast primers described in Table 1

[0016] cp1: forward sequence 5-gtggacctgactccttgaa-3, reverse sequence 5-agccgaggtcgtggtaa-3;

[0017] cp2: forward sequence 5-...

Images