| Rosa rugosa is the indigenously famous flower in China. It contains the aromatic, drought, and resistance to melasma and other good traits gene in its genome and is the germ plasm resource in Rosa L. plant breeding, while the short flowering period and monotonous flower color effect its application in gardens. Rosa rugosa and Rose chinensis hybridization is significantto improve raised germplasm, high entertaining and high disease resistance. However, the breeding processed slowly because the reason of two hybrid incompatibility was not clear for years.We intended to do interspecies cross between Rosa rugosa and Rose chinensis use amethod which throw a way of incising style to observe the change of growth on the top pollentube by Zhang S L. However, this method had difficult to draw a clear line of style form andpollen tube form so that the experimentation can’t carry on, and there needs a new method.In this research, the Rosa rugosa style extraction was used for the culture of Rosa rugosa and Rose chinensis pollen. The pollen tube top vesicles were marked with FM4-64and calciumin top of the pollen tube was marked with Fluo-3AM. The changes of the pollen tube morphological were observed by the optical microscope in order to study the influences of Rosa rugosa column extract on the pollen tube growth and analyze the cross-incompatibility mechanismbetween Rosa rugosa and Rose chinensis. The main research results were listed as follows:1. The changes of pollen germination rate, pollen tube length under different cultureconditions were obtained. The germination rate, pollen tube length of Rosa chinensis ‘RedCap’ pollen were all changed significantly after add Rosa rugosa ‘HC’ style extraction, whilethe change of Rosa rugosa ‘Zizhi’ has no decided change after through the same treatment.This result suggested that Rosa rugosa ‘HC’ style extraction can inhibit significantlygermination and pollen tube elongation of Rosa chinensis ‘Red Cap’, while the inhibition ofRosa rugosa ‘Zizhi’ is not significant.2. Observed by optical microscopy, the grow speed and top structure of Rosa chinensis‘Red Cap’ and Rosa rugosa ‘Zizhi’ which cultured in standard medium is normal. The pollentube growth speed of Rosa chinensis ‘Red Cap’ which cultured in Rosa rugosa ‘HC’ styleextraction liquid culture slowed down clearly, and part of pollen tube tip cell wall thicken,produced callose deposition, while the pollen tube growth speed of Rosa rugosa ‘Zizhi’ whichcultured in same culture slowed down slightly, growth form of it were normal, and there wereno callose deposition on it. 3. Under different culture conditions, tip vesicle distribution of pollen tube is changed. thepollen tube tip of Rosa chinensis ‘Red Cap’ and Rosa rugosa ‘Zizhi’ cultured in standardculture medium have a clearly V coloring, fluorescence was weakened gradually to subapical,it cultured in Rosa rugosa ‘HC’ column extraction liquid culture has also a clearly V coloringbut fluorescence intensity was weakened. In contrast, pollen tube tip fluorescence of Rosachinensis ‘Red Cap’ cultured in Rosa rugosa ‘Zizhi’ style extraction in liquid culture wasweak and vesicle distribution were no regularity. Generally speaking, there were a largenumber of vesicle aggregations on the normal growth rose pollen tube tip, and phenomenonof growth-slow, growth-stop, deformation appeared on no vesicle aggregation pollen tube.And in part of the pollen which are virtually invisible to vesicle distribution, pollen tube tipcell wall thickened, produced a large number of callose. This result indicated that Rose rugosastyle extract acted on pollen tube tip vesicles of Rose Chinensis, pollen tube growth stunted,and pollen tube tip produced callose deposition.4. Under different culture conditions, calcium ions distribution and calcium fluctuations ofpollen tube tip were different. Pollen tube top calcium ions of standard culture medium forculture of Rosa chinensis ‘Red Cap’ and Rosa rugosa ‘Zizhi’ cultured in Rosa rugosa ‘HC’style extraction liquid culture distribute concentrating at10-20μm, exists gradient change andthe calcium wave turbulence are significant. Calcium ions distribution gradient of Rosachinensis ‘Red Cap’ pollen tube tip which cultured in Rosa rugosa ‘HC’ column extractionliquid culture were not change obviously and calcium waves are weak, strong fluorescencewere observed in the morphology change site of pollen tube and two new splited pollen tubetip, indicated that calcium distribution is closely related to the growth of pollen tube.Generally speaking, Calcium ions distribution gradient of Rosa chinensis ‘Red Cap’ pollentube tip which cultured in Rosa rugosa ‘HC’ column extraction liquid culture disappeared,and pollen tube growth slowed down, deformated and stop eventually. Part of pollen tube tipcell wall thicken, produced callose deposition. This suggested that rose pollen tube tipcalcium ion distribution were destroyed by rose column extract, leading to pollen tube growthstunted, and pollen tube tip produced callose deposition. |
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