Development of a male specific genetic marker for Garcinia subelliptica Merr. tree

Figures & data

Figure 1. Flowers and fruits of the different sex types of G. subelliptica. (A) Male, (B) Female, (C) Hermaphrodite, (D) Fruits bearing in female tree, (E) Fallen fruit that produces an unpleasant smell

Table 1. Individuals of G. subelliptica used in this study

Sampling site	Estimated age of the tree	Number of plants		Geographic locality	Used for
		Male	Female
Yohena, Nago	70	16	16	26°39ʹ38.6”N 128°00ʹ53.6”E	RAD-seq analysis and validation of DNA markers
Yamashiro, Itoman	30	24	24	26°05ʹ02.7”N 127°41ʹ15.8”E	RAD-seq analysis and validation of DNA markers
Nago, Nago	20	40	35	26°37ʹ06.0”N 127°59ʹ01.6”E	Validation of DNA markers
Makabe, Itoman	20	21^a	20	26°06ʹ40.1”N 127°41ʹ11.4”E	Validation of DNA markers

^aA male tree bearing hermaphrodite flowers at some of its branches was exited.

Figure 2. The distribution of k-mers and the genome size estimation of G. subelliptica. (a) k-mer (k = 15 ~ 31) counts and their frequencies. (b) Estimated genome size based on the 15 ~ 31 k-mers

Figure 3. Results of PCR using primers: a primer set PMG1F1 and PMG1R1 designed from male-specific contig k141_82390. M: 1 kb Plus DNA Ladder (New England Biolabs), N: Negative control (H₂O)

Figure 4. Amplification pattern of FSDM locus in male and female individual. m: 1 kb Plus DNA Ladder (New England Biolabs); M: Male; F: Female; N: Negative control (H₂O)

Table 2. Sex identification using FSDM

Sexual phenotype^a	No. of plants tested	FSDM
		Present	Absent
Male	101	101	0
Female	95	0	95

^aSee the footnote on Table 1.

Table 3. Segregation of FSDM in progeny of 18 female trees

Name of female tree	No. of progeny seedlings	FSDM		Chi-square test^a	Location
		Present	Absent
Yaese No. 1	46	27	19	0.238	Gushikami, Yaese
Yaese No. 2	36	20	16	0.505	Gushikami, Yaese
Yaese No. 3	29	15	14	0.853	Gushikami, Yaese
Yaese No. 4	11	3	8	0.132	Gushikami, Yaese
Yaese No. 5	34	10	24	0.016	Gushikami, Yaese
Yaese No. 6	1	1	0	0.317	Gushikami, Yaese
Yaese No. 7	52	31	21	0.166	Gushikami, Yaese
Heiwa no mori No. 499	6	5	1	0.102	Yamashiro, Itoman
Heiwa no mori No. 530	10	3	7	0.206	Yamashiro, Itoman
Yagaji No. 158	3	2	1	0.564	Yagaji, Nago
Yagaji No. 198	15	3	12	0.020	Yagaji, Nago
Yagaji No. 221	14	8	6	0.593	Yagaji, Nago
Nago museum No. 1	4	1	3	0.317	Nago, Nago
Nago museum No. 2	1	0	1	0.317	Nago, Nago
Nago museum No. 3	31	17	14	0.590	Nago, Nago
Nago museum No. 6	22	14	8	0.201	Nago, Nago
Kumejima Nakazato No. 2	7	5	2	0.257	Maja, Kumejima
Kumejima Uezu No. 1	2	1	1	1.000	Nishime, Kumejima
Total	324	166	158	0.657

^aP-value. Chi-test was carried out with the expected segregation ratio 1:1.