Application of VNTR-specific Primers in Fertility Identification of Sugar Beet

doi:10.7668/hbnxb.20192531

Abstract

Abstract:

In order to find out the cytoplasm types of some sterile line and maintainer line materials in the process of sugar beet breeding, it provides a guiding direction for the next breeding work. Molecular marker primer TR1 was used to identify the molecular level of 30 test materials cytoplasm types, and to compare the fertility traits in the field. The results showed that the PCR electrophoresis bands of maintainer line 960767 and sterile line N9848 were 750,500 bp.We named these cytoplasm types as N1 and S,respectively.And 750 bp bands were detected by T152,T328,T334,T306,T360,as N1 cytoplasm.The 500 bp bands detected were T766,T154,N122,I-1,B-3,Z-1,41419,10320,B-3-23,S865,S301,S151,S327,S333,S115,S305,S153,S359,S137,S163,and S313,inferred to be S type cytoplasm.The materials for the bands detected between 500 bp and 750 bp were T302,and T116.Because T302 also had the ability to maintain the sterility of S301 in field investigations,T302 and T116 were classified into another type,N2 cytoplasm type.Some plants with 500, 750 bp of materials were T302, I-1 and B-3, which included both normal and Owen cytoplasm were characteristic materials. The results of field pollen fertility survey T766, T154, N122, I-1, B-3, Z-1, 414191, 10320 and B-3-23 were all fertile, but their cytoplasm type was identified to belong to S type cytoplasm. According to molecular identification and field fertility trait survey, the results of most materials were consistent with the results of fertility trait survey. Some materials had inconsistent molecular identification and fertility trait results, requiring further adjustment of the selection direction.

Key words: Sugar beet, Molecular markers, Fertility identification, TR1 primer

ZHANG Hui, WANG Liang, FU Zengjuan, LI Xiaodong, ZHAO Shangmin, E Yuanyuan, ZHENG Wenzhe, ZHANG Ziqiang, ZHANG Bizhou, ZHANG Huizhong. Application of VNTR-specific Primers in Fertility Identification of Sugar Beet[J]. Acta Agriculturae Boreali-Sinica, 2022, 37(2): 35-41. doi: 10.7668/hbnxb.20192531.

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Figures/Tables 8

Tab.1 Test materials names

序号 Number	材料名称 Material name	序号 Number	材料名称 Material name
1	960767	16	N9849
2	T766	17	S865
3	T302	18	S301
4	T152	19	S151
5	T328	20	S327
6	T334	21	S333
7	T116	22	S115
8	T306	23	S305
9	T154	24	S153
10	T360	25	S359
11	N122	26	S137
12	I-1	27	S163
13	B-3	28	S313
14	Z-1	29	10320
15	41419	30	B-3-23

Fig.1 1% agarose gel DNA detection
1—28.The electrophoresis detection results of the other test materials excluding 960767 and N9849.

Fig.2 Results of maintainer 960767 and sterile N9849 amplified by the TR1 primer PCR
1—8.The TR1 primer test results of different plants of maintainer material 960767;9—17.The TR1 primer test results of different plants of sterile line material N9849.

Fig.3 Detection results of some materials amplified by the TR1 primer PCR
1.T334;2.S333;4,5.S115;3,6.T116;8,10.T302;7,9.S301;11,12.960767;13—15.I-1;16—18.B-3;19—21.Z-1;22—24.41419;25—26.10320;27—28.B-3-23。

Fig.4 Detection results of maintainer material T302 and sterile material S301 amplified with the TR1 primer PCR
1—5.Detection of different plants of sterility material S301;6—10.Detection of different plants of sterile material S115;11—15.Detection of different plants of N2 cytoplasmic materials T302;16—20.Detection of different plants of N2 cytoplasmic materials T116.

Fig.5 Detection results of some fertile materials amplified by the TR1 primer PCR
1—6.Detection of different plants of 960767;7—13.Detection of different plants T152;14—20.Detection of different plants T328;21—27.Detection of different plants T334;28—31.Detection of different plants T306;32—35.Detection of different plants T360;36—40.Detection of different plants T302;41—44.Detection of different plants T116;45—48.Detection of different plants T766;49—54.Detection of different plants N122.

Fig.6 Test results of some sterile materials by TR1 primer PCR
1—2.Detection of different plants of S865;3—4.Detection of different plants of S301;5—6.Detection of different plants of S151;7—8.Detection of different plants of S327;9—10.Detection of different plants of S333;11—12.Detection of different plants of S115;13—14.Detection of different plants of S305;15—16.Detection of different plants of S315;17—18.Detection of different plants of S359;19—20.Detection of different plants of S137;21—22.Detection of different plants of S163;23—24.Detection of different plants of S313.

Tab.2 Identification of amplification and field breeding investigation by TR1 primers of different test materials

材料名称 Material name	条带位置/bp Band position	育性性状 Fertility character	类型 Type	材料名称 Material name	条带位置/bp Band position	育性性状 Fertility character	类型 Type
960767	750	可育型	N1	N9849	500	全不育型	S
T766	500	可育型	S	S865	500	半不育二型	S
T302	500—750	可育型	N2	S301	500	全不育型	S
T152	750	可育型	N1	S151	500	全不育型	S
T328	750	可育型	N1	S327	500	全不育型	S
T334	750	可育型	N1	S333	500	全不育型	S
T116	500—750	可育型	N2	S115	500	半不育二型	S
T306	750	半可育型	N1	S305	500	半不育一型	S
T154	500	半可育型	S	S153	500	半不育二型	S
T360	750/500	可育型	N1	S359	500	全不育型	S
N122	500	可育型	S	S137	500	半不育二型	S
I-1	500	可育型	S	S163	500	半不育二型	S
B-3	500	可育型	S	S313	500	全不育型	S
Z-1	500	可育型	S	10320	500	可育型	S
41419	500	可育型	S	B-3-23	500	可育型	S

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