Studies on Purification of Zizania latifolia Seedlings Based on the Detection of T-type Ustilago esculenta
PENG Hui1,*, YAO Liang-Hong2,*, ZHANG Zhen2, XIA Wen-Qiang1, LI Shi-Yu1, TANG Jin-Tian1, ZHANG Ya-Fen1, CUI Hai-Feng1, YE Zi-Hong1,**
1 College of Life Sciences/Zhejiang Provincial Key Laboratory of Biometrology and Inspection & Quarantine, China Jiliang University, Hangzhou 310018, China; 2 Wuzhen Agricultural Economic Service Center, Tongxiang 314501, China
Abstract:Jiaobai is an edible fleshy stem formed by the stem of Zizania latifolia infected by Ustilago esculenta. Gray Jiaobai often appears in field production, which seriously affects the yield and quality of Jiaobai. The infection and proliferation of T-type U. esculenta is the main factor causing gray Jiaobai, normal Jiaobai was formed by the infection of MT type strain, and long-term asexual breeding makes T-type strain and MT strain coexist in Z. latifolia seedlings. Therefore, it is very important for the development of Z. latifolia industry to purify the strains in Z. latifolia seedlings and eliminate T-type strain, so as to control the production of gray Jiaobai from the source. A specific detection method for T-type strain was established, and the detection limit was 0.008 8 ng/µL. Then, through the detection of the relative content of T-type strains in seedlings under different seedling raising methods, it is clear that the shell seedling of "with Jiaobai" seedling was the best way of initial purification, and the seedling raising in the upper part of bolt tube was the best way of seedling expansion. Based on the fact that the adaptability of T-type strain to stress was much higher than that of MT-type strain, finally, a small number of purified Z. latifolia seedlings without gray Jiaobai were obtained by 5 years stress treatment of Z. latifolia seedlings and the detection of T-type U. esculenta. The above results have theoretical and practical guiding significance for the breeding of Jiaobai seedlings.
彭辉, 姚良洪, 张真, 夏文强, 李士玉, 汤近天, 张雅芬, 崔海峰, 叶子弘. 基于 T 型菰黑粉菌检测的茭白种苗纯化研究[J]. 农业生物技术学报, 2023, 31(8): 1757-1766.
PENG Hui, YAO Liang-Hong, ZHANG Zhen, XIA Wen-Qiang, LI Shi-Yu, TANG Jin-Tian, ZHANG Ya-Fen, CUI Hai-Feng, YE Zi-Hong. Studies on Purification of Zizania latifolia Seedlings Based on the Detection of T-type Ustilago esculenta. 农业生物技术学报, 2023, 31(8): 1757-1766.
[1] 陈建明, 王来亮, 周锦连, 等. 2015. 浙江省茭白栽培新技术的探索与实践[J]. 长江蔬菜, 2015(22): 135-137. (Chen J M, Wang L L, Zhou J L, et al. 2015. Exploration and practice of new technology of Zizania latifolia cultivation in Zhejiang province[J]. Journal of Changjiang Veg-etables, 2015(22): 135-137.) [2] 陈可可, 杨新琴, 郑寨生, 等. 2022. 2 种留种育苗方法对茭白种苗质量、经济性状与产量的影响[J]. 浙江农业科学, 63(05): 17-19. (Chen K K, Yang X Q, Zheng Z S, et al. 2022. Effect of two seed methods on the quality, eco-nomic traits and yield of Zizania latifolia seedlings[J]. Journal of Zhejiang Agricultural Sciences, 63(05): 17-19.) [3] 陈可可, 张尚法, 葛鑫涛, 等. 2020. 双季茭白“带茭”苗二次留种育苗技术[J]. 长江蔬菜, 2020(24): 37-39. (Chen K K, Zhang S F, Ge X T, et al. 2020. Secondary seed retention and seedling raising technology of double cropping Zizania latifolia with gird[J]. Journal of Changjiang Veg-etables, 2020(24): 37-39.) [4] 丁小余, 徐祥生, 陈维培. 1991.“雄茭”、灰茭形成规律的初步研究[J]. 武汉植物学研究, 9(02): 115-120. (Ding X Y, Xu X S, Chen W P.1991. A preliminary study on the development of“male”and smutted tillers of Zizania caduciflora Hand. -Mazz[J]. Journal of Wuhan Botanical Research, 9(02): 115-120.) [5] 杜叶红.2019. 浙江省茭白不同栽培模式的效益分析[J]. 浙江农业科学, 60(03): 383-384, 417. (Du Y H.2019. Benefit analysis of Zizania latifolia under different culti-vation patterns in Zhejiang province[J]. Journal of Zheji-ang Agricultural Sciences, 60(03): 383-384, 417.) [6] 葛鑫涛.2020. 正常茭和灰茭特异性分子标记开发及其应用 [D]. 硕士学位论文, 中国计量大学. 导师: 张雅芬, 蒋明, pp. 9-10. (Ge X T.2020. Development and application of phenotype specific molecular markers of white Jiaobai and grey Jiaobai[D]. Thesis for M. S., China Jil-iang University, Supervisor: Zhang Y F, Jiang M, pp. 9-10.) [7] 葛鑫涛, 翁丽青, 郑春龙, 等. 2020. 双季茭白中菰黑粉菌遗传分化研究[J]. 植物生理学报, 56(04): 711-720. (Ge X T, Weng L Q, Zheng C L, et al. 2020. Study on genetic diversity of Ustilago esculenta in different double season varieties of Zizania latifolia[J]. Plant Physiology Jour-nal, 56(04): 711-720.) [8] 胡美华, 王来亮, 金昌林, 等. 2011. 单季茭白种苗繁育新技术 —— 薹管寄秧育苗法[J]. 长江蔬菜, 2011(23): 21-23. (Hu M H, Wang L L, Jin C L, et al. 2011. New tech-nology of seedling breeding of single - season Zizania latifolia - tube seedling breeding method[J]. Journal of Changjiang Vegetables, 2011(23): 21-23.) [9] 康璐瑶, 崔海峰, 张雅芬, 等. 2017. 不同茭白品种中菰黑粉菌的鉴定及 ISSR 多态性分析[J]. 菌物学报, 36(09):1210-1221. (Kang L Y, Cui H F, Zhang Y F, et al. 2017. Identification and ISSR polymorphism analysis of Usti- lago esculenta isolated from Zizania latifolia[J]. Myco-systema, 36(09): 1210-1221.) [10] 李智敏, 严理, 严准. 2016. 玉米瘤黑粉菌的寄生策略及其调控机制[J]. 微生物学报, 56(09): 1385-1397. (Li Z M, Yan L, Yan Z.2016. Parasitic strategy and regulation mechanism of Ustilago maydis - A review[J]. Acta Mi-crobiologica Sinica, 56(09): 1385-1397.) [11] 梁景玉.2007. 双季茭白的生物学特性及无公害优质栽培技术[J]. 上海蔬菜, 2007(01): 27-29. (Liang J Y.2007. Bi-ological characteristics and pollution-free high-quality cultivation techniques of double cropping Zizania latifo- lia[J]. Shanghai Vegetables, 2007(01): 27-29.) [12] 刘洪亮, 王险峰, 刘辉, 等. 2007. 玉米丝黑穗病原菌冬孢子萌发条件研究[J]. 中国植保导刊, 27(12): 5-8. (Liu H L, Wang X F, Liu H, et al. 2007. Study on germination conditions of teliospore of maize head smut[J]. China Plant Protection, 27(12): 5-8.) [13] 刘义满, 柯卫东, 黄新芳, 等. 2009. 茭白遗传多样性与育种技术[J]. 长江蔬菜, 2009(16): 1-4. (Liu Y M, Ke W D, Huang X F, et al. 2009. Genetic diversity and breeding techniques of Zizania latifolia[J]. Journal of Changjiang Vegetables, 2009(16): 1-4.) [14] 刘义满, 柯卫东, 李明华, 等. 2010. 茭白留种技术[J]. 种子,29(05): 112-113. (Liu Y M, Ke W D, Li M H, et al. 2010. Reserving seed technology of Zizania aquatica[J]. Seed, 29(05): 112-113.) [15] 陶忠富.2020. 浙江茭白种植现状及发展建议[J]. 中国果菜, 40(08): 73-76. (Tao Z F.2020. Planting situation and de-velopment suggestions of Zizania latifolia in Zhejiang province[J]. China Fruit&Vegetable, 40(08): 73-76.) [16] 王来亮, 陈建明, 马雅敏, 等. 2017. 利用茭白薹管结构差异快速辨别雄茭的方法[J]. 中国蔬菜, 2017(06): 47-49. (Wang L L, Chen J M, Ma Y M, et al. 2017. A new method for distinguishing unpregnant plants of Zizania latifolia based on difference among internal structure of culm[J]. China Vegetables, 2017(06): 47-49.) [17] 王树茂.2021. 雄茭和灰茭的发生原因及防治措施[J]. 安徽农学通报, 27(06): 48, 51. (Wang S M.2021. Causes and prevention measures of male and grey Zizania latifolia [J]. Anhui Agricultural Science Bulletin, 27(06): 48, 51.) [18] 谢贻格, 江扬先. 2019. 雄茭和灰茭的发生与防治[J]. 上海蔬菜, 2019(03): 24-25, 33. (Xie Y G, Jiang Y X.2019. Oc-currence and prevention of male and grey Zizania latifo-lia[J]. Shanghai Vegetables, 2019(03): 24-25, 33.) [19] 闫宁, 薛惠民, 石林豫, 等. 2013. 茭白“雄茭”和“灰茭”的形成及遗传特性[J]. 中国蔬菜, 2013(16): 35-42. (Yan N, Xue H M, Shi L Y, et al. 2013. The growth and heritabil-ity of Zizinia latifolia plants infected with sporidial strain of Ustilago esculenta[J]. China Vegetables, 2013 (16): 35-42.) [20] 尤文雨, 邹克琴, 俞晓平, 等. 2010. 菰黑粉菌的分离鉴定及其发酵液中植物激素的检测[J]. 菌物学报, 29(02):178-184. (You W Y, Zou K Q, Yu X P, et al. 2010. Isolation and identification of Ustilago esculenta and detection of plant hormones in the fermentation broth[J]. My-cosystema, 29(02): 178-184.) [21] 詹成.2011. 双季茭白施肥技术[J]. 上海蔬菜, 2011(04): 63. (Zhan C.2011. Fertilization technology of double crop-ping Zizania latifolia[J]. Shanghai Vegetables, 2011(04):63.) [22] 周杨.2016. 茭白不同育苗繁殖技术及其特点[J]. 浙江农业科学, 57(10): 1639-1641. (Zhou Y.2016. Different seed-ling propagation techniques and characteristics of Zizinia latifolia[J]. Journal of Zhejiang Agricultural Sci-ences, 57(10): 1639-1641.) [23] Chung K R, Tzeng D D.2004. Nutritional requirements of the edible gall-producing fungus Ustilago esculenta[J]. Jour-nal of Biological Sciences, 4(2): 246-252. [24] Guo H B, Li S M, Peng J, et al. 2007. Zizania latifolia Turcz. cultivated in China[J]. Genetic Resources and Crop Evo-lution, 54(6): 1211-1217. [25] Jose R C, Goyari S, Louis B, et al. 2016. Investigation on the biotrophic interaction of Ustilago esculenta on Zizania latifolia found in the Indo-Burma biodiversity hotspot[J]. Microbial Pathogenesis, 98: 6-15. [26] Kazuko Y, Moritaka S, Hisato Y, et al. 1996. Hypersensitivity pneumonitis induced by a smut fungus Ustilago esculenta [J]. Thorax, 51(6): 650-651. [27] Porebski S, Bailey L G, Baum B R.1997. Modification of a CTAB DNA extraction protocol for plants containing high polysaccharide and polyphenol components[J]. Plant Molecular Biology Reporter, 15(1): 8-15. [28] Swapan K S, Alak C D, Raju B.2016. Morphogenesis of edi-ble gall in Zizania latifolia (Griseb.) Turcz. ex Stapf due to Ustilago esculenta Henn. infection in India[J]. African Journal of Microbiology Research, 10(31): 1215-1223.Terrell E E, Batra L R. 1982. Zizania latifolia and Ustilago es-culenta, a grass-fungus association[J]. Economic Bota-ny, 36(3): 274-285. [29] Tu Z, Yamada S, Hu D, et al. 2019. Microbial diversity in the edible gall on white bamboo formed by the interaction between Ustilago esculenta and Zizania latifolia[J]. Cur-rent Microbiology, 76(7): 824-834. [30] Wang M, Zhao S, Zhu P, et al. 2018. Purification, characterization and immunomodulatory activity of water extract-able polysaccharides from the swollen culms of Zizania latifolia[J]. International Journal of Biological Macro-molecules, 107(Pt A): 882-890. [31] Wang M, Zhu P, Zhao S, et al. 2017. Characterization, antioxi-dant activity and immunomodulatory activity of polysac-charides from the swollen culms of Zizania latifolia[J]. International Journal of Biological Macromolecules, 95:809-817. [32] Wang Z H, Yan N, Luo X, et al. 2020. Gene expression in the smut fungus Ustilago esculenta governs swollen gall metamorphosis in Zizania latifolia[J]. Microbial Patho-genesis, 143: 104107. [33] Yan N, Du Y, Liu X, et al. 2018. Morphological characteris-tics, nutrients, and bioactive compounds of Zizania lati- folia, and health benefits of its seeds[J]. Molecules, 23(7): 1561. [34] Yang H C, Leu L S.1978. Formation and histopathology of galls induced by Ustilago esculenta in Zizania latifolia [J]. Phytopathology, 68(11): 1572-1576. [35] Ye Z H, Pan Y, Zhang Y F, et al. 2017. Comparative whole-ge-nome analysis reveals artificial selection effects on Usti- lago esculenta genome[J]. DNA Research, 24(6): 635-648. [36] Zhang J Z, Chu F Q, Guo D P, et al. 2012. Cytology and ultra-structure of interactions between Ustilago esculenta and Zizania latifolia[J]. Mycological Progress, 11(2): 499-508. [37] Zhang Y F, Cao Q C, Hu P, et al. 2017. Investigation on the differentiation of two Ustilago esculenta strains - impli-cations of a relationship with the host phenotypes ap-pearing in the fields[J]. BMC Microbiology, 17(1): 228. [38] Zhang Y F, Liu H L, Cao Q C, et al. 2018. Cloning and charac-terization of the UePrf1 gene in Ustilago esculenta[J]. Fems Microbiology Letters, 365(12): 4956762.