Pollination ecotypes and herkogamy variation of Hiptage benghalensis (Malpighiaceae) with mirror-image flowers

doi:10.17520/biods.2016248

Abstract

Abstract:

Herkogamy, the spatial separation of sexual organs within flowers, has long been thought to be a floral adaptation of reducing self pollination in hermaphrodites. Herkogamy of mirror-image flowers in the Asian endemic Hiptage benghalensis (Malpighiaceae) varies greatly between populations on Hainan Island (smaller than 4 mm) and Guizhou Province (larger than 10 mm). Such divergence in herkogamy could be adaptive to pollinator body size, which relates to pollen placement and stigma contact position. Recipient selection between local pollinators and plants may facilitate pollinator assembly in isolated populations, even forming pollination ecotypes. Pollinators were investigated in six geographic populations of H. benghalensis to study whether variations in herkogamy correspond to pollinator size. Molecular markers with nuclear ITS 1/4, chloroplast ycf1b and rbcL were also used to detect genetic differentiation between populations with large or small herkogamy. Our data showed small herkogamy in two populations from Hainan Island and one population from Xishuangbanna (southern Yunnan Province) as they were pollinated by small bees including Apis dorsata and Apis mellifera, with an abdomen width varying from 2 to 4 mm. Two populations from Guizhou Province and one population from Southeast Yunnan with larger herkogamy were pollinated by carpenter bees Xylocopa nasalis and bumble bees (Bombus sp.), with an obvious larger body size with a mean abdomen width of 11 mm. The style length (F = 12.5, P < 0.05) was also significantly different between large versus small herkogamous populations. Genetic data based on nuclear (ITS 1/4) and chloroplast (rbcL and ycf1) DNA also supported the differentiation between small and large herkogamous populations. These results indicated associations between flower and pollinator size, suggesting pollination ecotypes in H. benghalensis. This study indicated that the pollination ecotype in mirror-image flowers was associated with variations in herkogamy.

Key words: adaptive evolution, pollination efficiency, reproductive isolation, genetic distance, ycf1b, rbcL, ITS

Zhenna Qian, Qianwan Meng, Mingxun Ren. Pollination ecotypes and herkogamy variation of Hiptage benghalensis (Malpighiaceae) with mirror-image flowers[J]. Biodiv Sci, 2016, 24(12): 1364-1372.

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URL: https://www.biodiversity-science.net/EN/10.17520/biods.2016248

https://www.biodiversity-science.net/EN/Y2016/V24/I12/1364

Figures/Tables 6

Fig. 1 Mirror-image flowers and measurements of key floral traits in Hiptage benghalensis. R, Right-styled flower; L, Left-styled flower; HK, Herkogamy; SL, Style length; AS, Anther separation.

Table 1 Geographic locations and ratio of right- and left-styled flowers of Hiptage benghalensis populations

地点 Location	居群 Population	经纬度 Latitude/longitude	海拔 Altitude (m)	种群两种花型比率 Right/Left-styled flowers	χ ²	P
贵州 Guizhou	GZ-LK	25°05′24″ N, 106°12′59″ E	466	1:1 (21/21)	0	1
贵州 Guizhou	GZ-ZX	24°58′29″ N, 106°08′09″ E	442	1:1 (18/18)	0	1
云南 Yunnan	YN-YJ	23°20′17″ N, 102°11′52″ E	1,758	1:1.18 (11/13)	0.75	0.68
云南 Yunnan	YN-BN	21°52′16″ N, 101°19′27″ E	570	1:1 (32/32)	0	1
海南 Hainan	HN-BW	19°06′18″ N, 109°06′18″ E	223	1:1.50 (36/41)	1.2	0.27
海南 Hainan	HN-DL	18°43′19″ N, 109°52′24″ E	332	1:1.33 (28/39)	0.28	0.59

Table 2 DNA fragments and the primers used for genetic studies of Hiptage benghalensis populations

DNA片段	引物序列 Sequence	片段大小 Fragment size	参考文献 Reference
核基因 ITS 1/4	ITS1: TCCGTAGGTGAACCTGCG ITS4: TCCTCCGCTTATTGATATGC	651 bp	White et al, 1990
叶绿体 rbcL	1F: ATGTCACCACAAACAGAAAC 724R: TCGCATGTACCTGCAGTAGC	715 bp	Davis & Anderson, 2010
叶绿体 ycf1b	F: TCTCGACGAAAATCAGATTGTTGTGAAT R: ATACATGTCAAAGTGATGGAAAA	926 bp	Dong et al, 2015

Fig. 2 Boxplot of variations in herkogamy (A), style length (B) and anther separation (C) of mirror-image flowers among Hiptage benghalensis populations

Fig. 3 Pollinators in different geographic populations of Hiptage benghalensis. (A) and (B), Xylocopa nasalis visiting GZ-LK and GZ-ZX populations in Guizhou Province; (C) Bombus sp. visiting YN-YJ Population in Yunnan Province; (D) Pollen grains (red circles) on the thorax and abdomen of a carpenter bee; (E) Apis mellifera in YN-BN Population in Yunnan Province; (F) Apis dorsata in HN-BW Population on Hainan Island.

Fig. 4 Pollination ecotypes of Hiptage benghalensis. Neighbor-joining phylogenetic tree was drawn based on ITS1/4 and chloroplast (rbcLand ycf1b) DNA sequences. The histograms on the map refer to the mean values of herkogamy, and the unit of ordinate is millimeter (mm). ■ Herkogamy (righ-styled flower); □ Pollinator abdomen width; Herkogamy (left-styled flower).

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