Self-sterility May Be Due to Prezygotic Late-acting Self-incompatibility and Early-acting Inbreeding Depression in Chinese Chestnut

in Journal of the American Society for Horticultural Science

Chinese chestnut (Castanea mollissima), which is native to China, has been cultivated as a nontimber forest tree species for 4000 years. This species has been found to display self-sterility, which results in a significantly lower seed set following self-pollination (SP) compared with that following cross-pollination (CP). Self-sterility can be induced by prezygotic or postzygotic late-acting self-incompatibility (LSI) or early-acting inbreeding depression (EID). To elucidate the causes of self-sterility in chestnut, we investigated pollen–pistil interactions, fertilization, and early ovule development following SP and CP by using a paraffin section technique and fluorescence microscopy. The fruit set percentage and seed characteristics also were evaluated among different pollination treatments. The results show that there were no significant differences in pollen tube behavior following SP vs. CP, regardless of the stigmatic or stylar level. Double fertilization was significantly greater following CP (18.09%) than SP (2.58%). The significantly lower percentages of ovule penetration and double fertilization in the selfed vs. crossed ovules support a prezygotic LSI mechanism in C. mollissima. The fruit set resulting from chase-pollination (CHP; 53.85% to 63.64%) was greater than that resulting from SP (12.12% to 14.00%). In addition, the distribution of aborted seed sizes after SP showed a widely clumped pattern. Abortion occurred at different stages during seed development rather than at a uniform stage, which supported the idea that EID was operating in C. mollissima. Levels of self-sterility in the chinese chestnut trees ranged from 88.2% to 90.5%. Thus, partial prezygotic LSI and EID contributed to self-sterility in the C. mollissima ‘Yanshanzaofeng’, with prezygotic LSI rejecting part of the self-pollen in the ovary and EID aborting part of the self-fertilized seeds.

Contributor Notes

This study was supported by the National Natural Science Foundation of China (grant No. 31500554), Hunan Provincial Natural Science Foundation of China (grant No. 2018JJ3870), and ‘12th Five year’ Science and Technology Support Program of China (grant No. 2013BAD14B04).

Corresponding author. E-mail: zoufeng06@126.com.

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Article Figures

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    Pollen germination, pollen tube growth, and ovule penetration after self-pollination (SP) and cross-pollination (CP) in chinese chestnut: (A) a female flower of the ‘Yanshanzaofeng’, showing the stigma, style, and ovary; (B) pollen grain that had germinated on the stigma 6 d after SP; (C) pollen tubes that grew in the middle of the style 12 d after SP; (D) pollen tubes that had grown through the style 17 d after SP; (E) pollen grain that had begun to germinate on the stigma 3 d after CP; (F) pollen tubes that grew in the middle of the style 9 d after CP; (G) pollen tubes that had grown through the style 15 d after CP; (H) a pollen tube that had penetrated the ovule 20 d after SP; (I) a pollen tube that had penetrated the ovule 18 d after CP. Or = ovary; Ov = ovule; Pt = pollen tube; Sg = stigma; St = style.

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    Growth of selfed and crossed pollen tubes in carpels of chinese chestnut, shown by whether the most elongated pollen tube reached one-quarter, one-half, or three-quarters of the way down to the carpels or the lower end of the carpel.

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    Distorted pollen tubes grew in the ovary after self-pollination (SP) in chinese chestnut: (A) longitudinal section showing irregular pollen tubes that had reached the top of the ovary 18 d after SP; (B–C) longitudinal section showing degraded pollen tubes in the ovary 19 d after SP; (D–E) distorted pollen tube with a reversed and hampered tube in the nucellar tissue 20 d after SP; (F) swollen tip of a pollen tube that had failed to grow in the nucellar tissue 21 d after SP. Dpt = degraded pollen tube; Ov = ovule; Pt = pollen tube.

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    Normal double fertilization and early embryonic development after self-pollination (SP) and cross-pollination (CP) in chinese chestnut: (A) longitudinal section showing the inner and outer integuments and two polar nuclei located in the embryo sac 18 d after SP; (B) longitudinal section showing the inner and outer integuments and egg cells located in the embryo sac 18 d after SP; (C) longitudinal section showing the inner integument, polar nucleus, and two male nuclei located at the micropylar end 19 d after SP; (D) longitudinal section showing the inner integument and a male nucleus beginning to fuse with an egg cell in the embryo sac 20 d after SP; (E) longitudinal section showing the ovary wall, the inner and outer integuments, and a proembryo in the embryo sac 23 d after SP; (F) longitudinal section showing the inner and outer integuments and a large number of free endosperm nuclei forming a circular free nuclear layer along the embryo sac wall 25 d after SP; (G) transverse section showing the inner integument and a male nucleus beginning to fuse with an egg cell in the embryo sac 18 d after CP; (H) longitudinal section showing the inner and outer integuments, free endosperm nuclei and a proembryo in the embryo sac 21 d after CP; (I) longitudinal section showing free endosperm nuclei in the embryo sac 22 d after CP; (J) longitudinal section showing the inner and outer integuments and a large number of free endosperm nuclei forming a circular free nuclear layer along the embryo sac wall 23 d after CP; (K) longitudinal section showing a globular embryo 30 d after SP; (L) longitudinal section showing free endosperm nuclei and a globular embryo 27 d after CP. Ec = egg cell; Es = embryo sac; Fen = free endosperm nuclei; Ge = globular embryo; Ii = inner integument; Mn = male nucleus; Ow = ovary wall; Oi = outer integument; Pe = proembryo; Pn = polar nuclei.

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    Degenerating and aborted ovules in self-pollinated (SP) ovaries at various stages in chinese chestnut: (A) longitudinal sections showing that the aborted ovules were darkly stained 20 d after SP; (B) longitudinal section showing aborted ovules with shrunken embryo sacs 21 d after SP; (C) longitudinal section showing degenerated ovules with cavities in the embryo sacs 22 d after SP; (D) longitudinal section showing aborted ovules with degenerated postament 23 d after SP; (E−F) longitudinal section showing aborted ovules 25 d after SP. Ao = aborted ovule; Do = degenerate ovule; Ii = inner integument; Ow = ovary wall; Oi = outer integument.

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    Mature fruit, mature seeds, and aborted seeds after self-pollination (SP), cross-pollination (CP), chase-pollination (CHP), and nonpollination (NP) treatments in chinese chestnut: (A) mature cupula and seed characteristics following CP; (B) mature cupula and seed characteristics following SP; (C) mature cupula and aborted seed characteristics following CHP; (D) mature cupula and aborted seed characteristics following NP; (E) comparison of seed characteristics between the NP and SP treatments; (F) mature seed characteristics following CP and CHP. Cps = CP seed; Sps = SP seed; Sss = self-sterile seed.

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    Distributions of aborted seed length and the cv for chinese chestnut in the self- and cross-pollination treatments.

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