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The in vitro germination and storage characteristics of Keteleeria fortunei var. cyclolepis pollen provide a reference for cross breeding

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Abstract

Keteleeria fortunei var. cyclolepis is an ideal tree species for mountain afforestation, timber forests, and landscaping. Its pollination process can be affected by the rainy season, making it difficult to pollinate the massive female cones, which leads to a high abortion rate and low quality of seeds. Here, we observed the pollen morphology of K. f. cyclolepis using scanning electron and light microscopes, investigated the characteristics of its in vitro germination by the detached method, and explored the effect of different storage temperatures and times on the pollen germination rate and the activity of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT). Our results indicated that the pollen of K. f. cyclolepis is a five-cell pollen, comprising one noumenon and two air sacs, both of which were oval in polar view. The optimal condition for pollen germination of K. f. cyclolepis was 240 g/L sucrose + 70 mg/L CaCl2 + 210 mg/L H3BO3 at 24 °C and pH 6.0, resulting in a germination rate of 45.0%. The effects of different storage temperature and time on pollen germination rate varied significantly. The best storage temperature was − 80 °C, at which the germination rate was 20.9% after 365 days of storage, and the activity of three protective enzymes remained relatively high, representing relatively strong antioxidation and antiaging activity. Stepwise regression analysis showed that SOD was the main factor affecting the pollen germination rate of K. f. cyclolepis. The function of the three protective enzymes differed under various temperatures, for example, SOD served as a sensitive protective enzyme at room temperature, − 20 °C and − 80 °C, whereas both SOD and CAT served as sensitive protective enzymes at 4 °C.

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References

  • Abdullah YAM, Abul HMM, Obaidul IM, Ali M (2000) Cross compatibility between Abelmoschus esculentus and A. moschatus. Euphytica 114:175–180

    Google Scholar 

  • Bilderback DE (1981) Impatiens pollen germination and tube growth as a bioassay for toxic substances. Environ Health Persp 37:95–103

    CAS  Google Scholar 

  • Dong B, Zheng X, Liu H, Able JA, Yang H, Zhao H, Zhang M, Qiao Y, Wang Y, Liu M (2017) Effects of drought stress on pollen sterility, grain yield, abscisic acid and protective enzymes in two winter wheat cultivars. Front Plant Sci 8:1008

    PubMed  PubMed Central  Google Scholar 

  • Doyle JH, Verhoeven RL, Bester C, Wingfield BD, Botha AM, Bornman CH (2002) Germ-furrow morphology and storage conditions determine the degree of viability of Pinus caribaea pollen. S Afr J Bot 68:457–463

    Google Scholar 

  • Fecht-Christoffers MM, Fuhrs H, Braun HP, Horst WJ (2006) The role of hydrogen peroxide-producing and hydrogen peroxide-consuming peroxidases in the leaf apoplast of cowpea in manganese tolerance. Plant Physiol 140:1451–1463

    CAS  PubMed  PubMed Central  Google Scholar 

  • Fei S, Nelson E (2003) Estimation of pollen viability, shedding pattern, and longevity of Creeping bentgrass on artificial media. Crop Sci 43:2177–2181

    Google Scholar 

  • Flores-Rentería L, Whipple AV, Benally GJ, Patterson A, Canyon B, Gehring CA (2018) Higher temperature at lower elevation sites fails to promote acclimation or adaptation to heat stress during pollen germination. Front Plant Sci 9:536

    PubMed  PubMed Central  Google Scholar 

  • Fragallah SADA, Lin S, Li N, Ligate EJ, Chen Y (2019) Effects of sucrose, boric acid, pH, and incubation time on in vitro germination of pollen and tube growth of Chinese fir (Cunnighamial lanceolata L.). forests 10:102

  • Guo Y, Zhao Y (2018) Effects of storage temperature on the physiological characteristics and vegetative propagation of desiccation-tolerant mosses. Biogeosciences 15:1–16

    Google Scholar 

  • He YH, Jiang Y, Huang RL, Liu F, Jiang Y (2017) Differences of seed quality in Keteleeria cyclolepis flous from different provenances. J Cent South Univ For Technol 37:38–41

    Google Scholar 

  • Hirose T, Hashida Y, Aoki N, Okamura M, Yonekura M, Ohto C, Terao T, Ohsugi R (2014) Analysis of gene-disruption mutants of a sucrose phosphate synthase gene in rice, OsSPS1, shows the importance of sucrose synthesis in pollen germination. Plant Sci 225:102–106

    CAS  PubMed  Google Scholar 

  • Horsley TN, Johnson SD, Stanger TK (2007) Optimising storage and in vitro germination of Eucalyptus pollen. Aust J Bot 55:83–89

    CAS  Google Scholar 

  • Huang RL, Jiang Y, Liu F, Liu XS, Jiang Y (2018) A study on genetic variations of seeding traits of plus trees of Keteleeria fortune var. cyclolepis. For Sci Technol 12:3–7

    Google Scholar 

  • Jia WQ, Wang SP, Li LY (2015) Pollen morphology, storage condition and physiologically dynamic change during storage of Camellia magniflora. Acta Bot Boreal –Occident Sin 35:754–760

    CAS  Google Scholar 

  • Jones LH (1983) Experiments in plant tissue culture. Trends in Biochem Sci 8:343

    Google Scholar 

  • Kamienska A, Durley RC, Pharis RP (1976) Isolation of gibberellins A3, A4 and A7 from Pinus attenuata pollen. Phytochemistry 15:421–424

    CAS  Google Scholar 

  • Kanazawa S, Sano S, Koshiba T, Ushimaru T (2000) Changes in antioxidative enzymes in cucumber cotyledons during natural senescence: comparison with those during dark-induced senescences. Physiol Plantarum 109:211–216

    CAS  Google Scholar 

  • Kaur D, Singhal VK (2019) Meiotic abnormalities affect genetic constitution and pollen viability in dicots from Indian cold deserts. BMC Plant Biol 19:10

    PubMed  PubMed Central  Google Scholar 

  • Kitoh T, Matsushita M (1980) A new staining method of astrocytes for paraffin section. Acta Neuropathol 49:67–69

    CAS  PubMed  Google Scholar 

  • Li HS (2000) Experimental principles and techniques of plant. Physiology and biochemistry. Higher Education Press, Beijing

    Google Scholar 

  • Li GP, Huang QC (2006) Ontogeny of pollen and pollination in Keteleeria fortune. Sci Silva Sin 5:42–47

    Google Scholar 

  • Li GP, Huang QC, Yang LS, Qin GY (2007) In vitro pollen germination and pollen tube growth of Pinus thunbergii. Forest Res 20:224–229

    CAS  Google Scholar 

  • Lin Y, Wang Y, Iqbal A, Shi P, Li J, Yang Y, Lei X (2017) Optimization of culture medium and temperature for the in vitro germination of oil palm pollen. Sci Hortic 220:134–138

    CAS  Google Scholar 

  • Liu L, Huang L, Li Y (2013a) Influence of boric acid and sucrose on the germination and growth of areca pollen. Am J Plant Sci 4:1669–1674

    CAS  Google Scholar 

  • Liu YP, Zhu YL, Ma YT, Zhang JT, Zhai XQ, Luo XY (2013b) Effects of different storage conditions on germination rate and protective enzymes activity of Magnolia liliflora Desr. pollen. J Northeast For Univ 41:59–61

    Google Scholar 

  • Liu XS, Jiang Y, Huang RL, Liu F, Xiao YF, Jiang Y, Wei SX (2017) Variation in traits of cone and seed of Keteleeria fortunei var. cyclolepis and its relationship with environmental factors. Guihaia 37:118–126

    Google Scholar 

  • Mao P, Zhang Y, Cao B, Guo L, Shao H, Cao Z, Jiang Q, Wang X (2016) Effects of salt stress on eco-physiological characteristics in Robinia pseudoacacia based on salt-soil rhizosphere. Sci Total Environ 568:118–123

    CAS  PubMed  Google Scholar 

  • Mercado JA, Fernfindez-Mufiozb A, Quesada MA (1994) In vitro germination of pepper pollen in liquid medium. Sci Hortic-amsterdam 57:273–281

    Google Scholar 

  • Monteiro D (2005) Phosphoinositides and phosphatidic acid regulate pollen tube growth and reorientation through modulation of [Ca2+] c and membrane secretion. J Exp Bot 56:1665–1674

    CAS  PubMed  Google Scholar 

  • Murakami Y, Yamaguchi S (2010) Distinctive traits of Camellia vernalis, C. sasanqua and C. japonica detected by the interaction of pollen grains in mixed pollen culture. Breeding Sci 49:131–133

    Google Scholar 

  • Nygaard P (2010) Studies on the germination of pine pollen (Pinus mugo) in vitro: III. Inhibition by D-mannose and deoxyhexoses. Physiol Plantarum 23:372–384

    Google Scholar 

  • Raohavan V, Baruah HK (2010) Effect of time factor on the stimulation of pollen germination and pollen tube growth hy certain auxinh, vitamins, and trace elements. Physiol Plantarum 12:441–451

    Google Scholar 

  • Rodriguez-Enriquez MJ, Mehdi S, Dickinson HG, Grant-Downton RT (2013) A novel method for efficient in vitro germination and tube growth of Arabidopsis thaliana pollen. New Phytol 197:668–679

    CAS  PubMed  Google Scholar 

  • Shivanna KR, Johri BM (1985) The angiosperm pollen: structure and function. Wiley Eastern, New Delhi

    Google Scholar 

  • Smith GF, Tiedt LR, Rapid A (1991) Non-destructive osmium tetroxide technique for preparing pollen for scanning electron microscopy. Taxon 40:195–200

    Google Scholar 

  • Tan JH (2011) Effects of storage temperature and storage time on germination rate and protective enzymes activity of Pinus massoniana pollen. Sci Silva Sin 47:28–32

    Google Scholar 

  • Tian J, Shang FN, Song SY, Wang J, Kang XY (2012) In vitro pollen germination and characteristics on pollen tube of Pinus tabulaeformis Carr. Chinese Agric Sci Bull 28:21–27

    Google Scholar 

  • Wang Q, Lu L, Wu X, Li Y, Lin J (2003) Boron influences pollen germination and pollen tube growth in Picea meyeri. Tree Physiol 23:345–351

    CAS  PubMed  Google Scholar 

  • Wolukau JN, Zhang S, Xu G, Chen D (2004) The effect of temperature, polyamines and polyamine synthesis inhibitor on in vitro pollen germination and pollen tube growth of Prunus mume. Sci Hortic 99:289–299

    CAS  Google Scholar 

  • Zhang JT (1989) Studies on the pollen morphology of Pinaceae in China. Bull Bot Res 3:87–98

    Google Scholar 

  • Zhang G, Tanakamaru K, Abe J, Morita S (2007) Influence of waterlogging on some anti-oxidative enzymatic activities of two barley genotypes differing in anoxia tolerance. Acta Physiol Plant 29:171–176

    Google Scholar 

  • Zhao LJ, Li SJ, Yu JH, Wang BC, Yang CP (2011) In vitro germination and cytology analysis of Populus simonii × P. nigra pollen. Sci Silva Sin 47:36–41

    Google Scholar 

  • Zou Q (2000) Instruction for plant physiology experiments. China Agricultural Publishing House, Beijing

    Google Scholar 

Download references

Funding

This study was supported by the Independent Research Fund of the Guangxi Key Laboratory of Superior Timber Trees Resource Cultivation (No. 2019-A-03-01) and Guangxi Key Research and Development Plan (No. AB163800100).

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Authors and Affiliations

  1. Guangxi Key Laboratory of Superior Trees Resource Cultivation, Guangxi Zhuang Autonomous Region Forestry Research Institute, Nanning, 530002, China

    Xiongsheng Liu, Yufei Xiao, Yong Wang, Fengfan Chen, Ronglin Huang & Yi Jiang

Authors
  1. Xiongsheng Liu
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  2. Yufei Xiao
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  3. Yong Wang
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  4. Fengfan Chen
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  5. Ronglin Huang
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  6. Yi Jiang
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Contributions

All authors contributed to the study conception and design. Xiongsheng Liu, Yufei Xiao, and Yi Jiang designed the experiments and wrote the paper; Yong Wang, Fengfan Chen, and Ronglin Huang participated in the experiment, helped to complete the experiments and analyzed the data; Yi Jiang revised the article, and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript. Xiongsheng Liu and Yufei Xiao contributed equally to this work.

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Correspondence to Yi Jiang.

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Liu, X., Xiao, Y., Wang, Y. et al. The in vitro germination and storage characteristics of Keteleeria fortunei var. cyclolepis pollen provide a reference for cross breeding. Protoplasma 257, 1221–1230 (2020). https://doi.org/10.1007/s00709-020-01509-w

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  • DOI: https://doi.org/10.1007/s00709-020-01509-w

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