Genetic Mapping for a Medium Short-fruit Mutant of Cucumber

doi:10.16420/j.issn.0513-353x.2021-0194

Acta Horticulturae Sinica ›› 2021, Vol. 48 ›› Issue (7): 1359-1370.doi: 10.16420/j.issn.0513-353x.2021-0194

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Genetic Mapping for a Medium Short-fruit Mutant of Cucumber

CHENG Feng, SONG Mengfei, CAO Lei, ZHANG Mengru, YANG Zhige, CHEN Jinfeng, LOU Qunfeng^*()

State Key Laboratory of Crop Genetics and Germplasm Enhancement,College of Horticulture,Nanjing Agricultural University,Nanjing 210095,China

Received:2021-03-29 Revised:2021-05-06 Online:2021-07-25 Published:2021-08-10
Contact: LOU Qunfeng E-mail:qflou@njau.edu.cn

Abstract

Abstract:

We isolated a natural mutant,msf（medium short-fruit）,from the cucumber inbred line CCMC,a North China type with long fruits. The mutant showed the phenotype of shorter fruit length and fruit neck length than the wild type and also showed short plant,abnormal development of lateral branches and fragile main stem. Through paraffin section and scanning electron microscope observation,it was found that the difference of mutant fruit length was caused by the change of cell size. Genetic analysis showed that the segregation ratio of wild-type phenotype long fruit and mutant phenotype medium short fruit in F₂ segregation population conformed to Mendel’s 3︰1 genetic law. This indicated that msf mutation was a recessive trait controlled by a single gene. BSA-seq analysis showed that the candidate gene was located in a 26.7-30.9 Mb region of chromosome 1 of cucumber genome. The msf gene was mapped to a 28.4-29.8 Mb region using a F₂ population containing 437 individuals from a cross between msf and ‘hazerd’（a European greenhouse-type inbred line with short fruit）. CsaV3_1G044310 was identified as the candidate gene by genome-wide screening combined with dCAPS marker verification,which encodes a homologous protein of Arabidopsis type Ⅱ inositol polyphosphate 5-phosphatase（type Ⅱ 5 Ptase）. The quantitative expression of gene and the determination of endogenous hormones auxin and gibberellin the mutant may cause the change of hormone levels and affect the growth and development of plant.

Key words: cucumber, fruit length, mutant, gene mapping, type Ⅱ inositol polyphosphate 5-phosphatase;

CLC Number:

S642.2

CHENG Feng, SONG Mengfei, CAO Lei, ZHANG Mengru, YANG Zhige, CHEN Jinfeng, LOU Qunfeng. Genetic Mapping for a Medium Short-fruit Mutant of Cucumber[J]. Acta Horticulturae Sinica, 2021, 48(7): 1359-1370.

Figures/Tables 10

Fig. 1 Phenotypic characteristics of wild type and msf mutant

Fig. 2 Fruit length and width of‘Changchun Mici’wild type and msf mutant

Fig. 3 Observation of paraffin sections of fruit cell size in wild type（WT）and mutant（msf）at different stages after flowering “**”indicates significant difference from WT at the 0.01 probability level. Scale bars represent 20 μm.

Fig. 4 Scanning electron microscope observation of epidermis cells and pulp cells in wild type（WT）and mutant（msf）fruit at 16 d after flowering

Table 1 Phenotypic data for short fruit mutant and wild-type cucumber

群体	总株数	长果株数	中短果株数	期望比	χ²	P
Population	Total	Long fruit	Medium Short fruits	Expected ratio	χ²	P
P1（WT）	20	20	0
P2（msf）	20	0	20
F₁	40	40	0
F₂	447	339	108	3︰1	0.17	0.68

Fig. 5 Mapping of the msf gene A：Distribution of ∆（All-index）in two pools on seven chromosomes.（Each point represents a SNP site,and different chromosomes are distinguished by different colors;the black line is the SNP-index mean line,the blue line is the 95% threshold line,and the purple line is the 99% threshold line）;B：Distribution of ∆（All-index）in two pools on chromosome 1.（Each point represents a SNP site,the red line is the SNP-index mean line,the green line is the 95% threshold line, and the purple line is the 99% threshold line）;C：The msf gene was mapped to a region on chromosome 1 in an F2 population containing 437 individuals from a cross between msf and‘hazerd’.

Table 2 Candidate loci in the target region on chromosome 1

位置/bp Position	参考基因组 REF	碱基Base		SNP指数 SNP-index		类型 Category	氨基酸变化 Amino acid change
		野生型	变异	野生池	突变池
		WT	ALT	WT-pool	msf-pool
28 730 253	T	T	A	0.28	0.96	基因间Intergenic
28 790 458	A	A	T	0.28	0.91	上游 Upstream
29 026 627	G	G	C	0.25	1	基因间Intergenic
29 045 423	G	G	A	0.23	0.93	基因间Intergenic
29 057 944	C	C	T	0.21	0.96	基因间Intergenic
29 114 093	T	T	G	0.29	0.97	基因间 Intergenic
29 380 231	G	G	A	0.26	0.92	基因间 Intergenic
29 440 371	C	C	G	0.25	0.9	内含子Intronic
29 501 452	G	G	A	0.19	1	内含子Intronic
29 567 774	G	G	C	0.27	1	外显子Exonic	谷氨酰胺到谷氨酸 Q to E
29 647 321	G	G	A	0.26	0.9	基因间Intergenic
29 765 691	G	G	A	0.25	0.96	内含子Intronic
29 825 348	A	A	C	0.24	0.95	基因间Intergenic

Fig. 6 Relative expression level of CsaV3_1G044310 in different tissues of WT and msf plant

Fig. 7 Candidate gene structure（A）and protein domain（B） The arrow is the position of mutation.

Fig. 8 Endogenous IAA and GA content in different tissues of WT and mutant msf plants

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Genetic Mapping for a Medium Short-fruit Mutant of Cucumber

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