Optimized Culture Medium and Effect of Piriformospora indica on Growth of Rice Plants
-
摘要:
目的 内生真菌印度梨形孢Piriformospora indica（Pi）可定殖在多种植物根系中，促进植物生长发育，增强植物抗逆性。 方法 将印度梨形孢接种至8种供试培养基中，测定其在8种培养基中的生长速率，并将印度梨形孢与水稻幼苗共培养，分析印度梨形孢对水稻幼苗株高、叶长、根长、根数、鲜重和叶绿素含量等生物学性状的影响。 结果 8种供试培养基中，V8培养基是最适合印度梨形孢生长的培养基，且加入一定量蔗糖能进一步促进印度梨形孢的生长；研究还发现，接种了印度梨形孢的水稻株高、叶长、根长、根数、叶绿素含量和地上地下部鲜质量都显著高于对照；与对照相比，接种印度梨形孢20 d后的水稻，其株高增高了32.36%，叶绿素含量提高了15.88%。 结论 印度梨形孢是通过增加光合作用和增强根系生理机能，从而促进水稻的生长发育。 Abstract:Objective Enhancement on the growth, development and stress resistance of rice plants from the colonization of the endophytic fungus Piriformospora indica (Pi) on the roots was investigated. Method Pi was inoculated on 8 different media for optimal culture and co-cultured on rice seedlings to determine the biological improvements on the plants. Result Medium V8 was found to be most desirable for Pi growth. Addition of a certain amount of sucrose would further increase the effect. The plant height, leaf length, root length, root count, chlorophyll content and quality of the above- and underground parts of the treated rice plants were significantly greater than those of control. Notably, 20 days after the inoculation, the plant height increased by 32.36% and chlorophyll content 15.88%. Conclusion Pi significantly promoted the growth and development of rice plants as it symbiotically enhanced the photosynthesis and root physiology of the host. -
Key words:
- Piriformospora indica /
- medium /
- rice /
- growth enhancement
-
图 1 印度梨形孢在不同固体培养基上培养5、7和10 d时的形态
注：A为V8；B为农夫果园芒果；C为农夫果园橙子；D为农夫果园番茄；E为PDA；F为Aspergillus；G为MS；H为康乃馨。图 2同。
Figure 1. Morphology of Pi cultured on solid media for 5 d, 7 d and 10 d
Note:A:V8;B:Farmer orchard mango; C:Farmer orchard orange; D:Farmer orchard tomato; E:PDA; F:Aspergillus; G:MS; H:Carnation.Same for Fig. 2.
图 2 印度梨形孢在不同液体培养基中培养7 d时的形态
注：A为V8；B为农夫果园芒果；C为农夫果园橙子；D为农夫果园番茄；E为PDA；F为Aspergillus；G为MS；H为康乃馨。
Figure 2. Morphology of Pi in suspension cultures for 7 d
Note:A:V8;B:Farmer orchard mango; C:Farmer orchard orange; D:Farmer orchard tomato; E:PDA; F:Aspergillus; G:MS; H:Carnation.
图 3 印度梨形孢在含不同碳源的V8培养基上培养3 d、5 d和7 d时的形态
注：A：V8+1%蔗糖；B：V8；C：V8+1%葡萄糖；D：V8+1%果糖。
Figure 3. Morphology of Pi cultured on Medium V8 with varied carbon sources for 5 d, 7 d and 10 d
Note:A:Medium V8 with 1% sucrose; B:V8;C:Medium V8 with 1% glucose; D:Medium V8 with 1% fructose.
图 5 印度梨形孢对水稻幼苗生长的影响
注:A为水稻种子萌发7 d；B为水稻幼苗与Pi共培养7 d；C为水稻幼苗与Pi共培养20 d。
Figure 5. Effect of Pi on growth of rice seedlings
Note:A:Rice seedlings 7 d after seed germination; B:Rice seedlings co-cultured with Pi for 7 d; C:Rice seedlings co-cultured with Pi for 20 d.
表 1 印度梨形孢在不同固体培养基上的生长速率
Table 1. Growth rates of Pi on solid media
培养基
Medium纯生长量Net growth /cm 5 d 7 d 10 d V8 7.32±0.065 7.50 7.50 农夫果园芒果Farmer orchard mango 7.01±0.047 7.50 7.50 农夫果园橙子Farmer orchard orange 7.00±0.044 7.50 7.50 农夫果园番茄Farmer orchard tomato 6.92±0.087 7.50 7.50 PDA 3.03±0.097 4.70±0.079 7.38±0.044 Aspergillus 3.12±0.055 4.15±0.081 6.12±0.051 MS 2.51±0.117 3.97±0.101 5.57±0.047 康乃馨Carnation 2.00±0.036 3.22±0.051 4.69±0.035 
下载: 导出CSV
表 2 印度梨形孢在不同液体培养基中的质量变化
Table 2. Weight of Pi in suspension media
培养基
Medium5 d 7 d 10 d 鲜重
Fresh weight/g干重
Dry weight/g鲜重
Fresh weight/g干重
Dry weight/g鲜重
Fresh weight/g干重
Dry weight/gV8 3.931±0.121 0.776±0.013 3.034±0.163 0.788±0.007 2.412±0.035 0.797±0.005 Aspergillus 2.410±0.036 0.480±0.007 3.909±0.048 0.776±0.009 2.909±0.082 0.792±0.007 PDA 2.319±0.035 0.463±0.007 3.662±0.064 0.732±0.012 3.224±0.036 0.795±0.007 农夫果园番茄Farmer orchard tomato 2.236±0.015 0.447±0.005 3.595±0.039 0.717±0.006 3.334±0.049 0.794±0.005 农夫果园橙子Farmer orchard orange 1.298±0.013 0.253±0.004 2.704±0.014 0.537±0.004 3.945±0.048 0.790±0.008 农夫果园芒果Farmer orchard mango 0.459±0.009 0.089±0.002 0.893±0.008 0.179±0.004 2.098±0.021 0.407±0.005 MS 0.343±0.007 0.067±0.001 0.616±0.006 0.121±0.003 1.490±0.014 0.295±0.002
下载: 导出CSV
表 3 印度梨形孢在含不同碳源的V8培养基上的生长速率
Table 3. Growth rates of Pi on Medium V8 with varied carbon sources
培养基
Medium纯生长量Net growth /cm 5 d 7 d 10 d V8 3.23±0.09 5.87±0.17 7.19±0.06 V8+蔗糖V8+Sucrose 3.89±0.14** 6.46±0.16** 7.39±0.04** V8+葡萄糖V8+Glucose 2.89±0.12* 4.95±0.23** 6.61±0.27* V8+果糖V8+ Fructose 2.94±0.10* 4.98±0.18** 6.59±0.23* 注：*或**分别表示加糖和不加糖测定的数据间差异达到显著或极显著水平(T.TEST)。
Note: * or ** indicates statistically significant or extremely significant differences between sugar treatment and control (t-test).
下载: 导出CSV
表 4 印度梨形孢对水稻幼苗生物学性状的影响
Table 4. Effect of Pi on biological properties of co-cultured rice
生物学性状
Biological characteristicsP. indica处理
P. indica treatment对照
Control株高Plant height /cm 13.54±0.89** 10.23±0.74 最大叶长Maximum leaf length /cm 8.39±0.7 ** 6.67±0.82 最大叶宽Maximum leaf width /cm 2.39±0.26 2.34±0.33 根长Root length /cm 5.51±0.83** 4.65±0.56 根数Number of roots 10.3±1.25* 9.3±1.06 根鲜重Root fresh weight/g 0.019±0.002* 0.016±0.002 地上部鲜重Aboveground fresh weight/g 0.049±0.003 ** 0.038±0.005 叶绿素含量
Chlorophyll content /(mg·g-1)6.35±0.21** 5.48±0.33 注：*或**分别表示Pi处理组和对照组测定的指标间差异达到显著或极显著水平(T.TEST)。
Note: * or ** indicates statistically significant or extremely significant differences between Pi processing and control data(T.TEST).
下载: 导出CSV
-
[1] GNANAMANICKAM S S. Rice and its importance to human life[C]//GNANAMANICKAM S S. Biological Control of Rice Diseases. Berlin: Springer-Verlag, 2009: 1-11. doi: 10.1007%2F978-90-481-2465-7_1 [2] COUCH B C, KOHN L M. A multilocus gene genealogy concordant with host preference indicates segregation of a new species, Magnaporthe oryzae, from M. grisea[J]. Mycologia, 2002, 94(4):683-693. doi: 10.1080/15572536.2003.11833196 [3] 任小平, 谢关林, 赵丽涵.水稻纹枯病拮抗细菌的筛选与利用[J].植物保护学报, 2005, 32(4):337-342. doi: 10.3321/j.issn:0577-7518.2005.04.001REN X P, XIE G L, ZHAO L H. Screening and utilization of antagonistic bacteriae against rice sheath blight[J]. Journal of Plant Protection, 2005, 32(4):337-342.(in Chinese) doi: 10.3321/j.issn:0577-7518.2005.04.001 [4] VERMA S, VARMA A, REXER K H, et al. Piriformospora indica, gen. et sp. nov., a new root-colonizing fungus[J]. Mycologia, 1998, 90(5):896-903. doi: 10.1080/00275514.1998.12026983 [5] ANSARI M W, TRIVEDI D K, SAHOO R K, et al. A critical review on fungi mediated plant responses with special emphasis to Piriformospora indica, on improved production and protection of crops[J]. Plant Physiology & Biochemistry, 2013, 70(1):403-410. http://www.wanfangdata.com.cn/details/detail.do?_type=perio&id=e4a971e703e8cca347df1b17d002efe5 [6] SINGH A, SHARMA J, REXER K H, et al. Plant productivity determinants beyond minerals, water and light:Piriformospora indica, a revolutionary plant growth promoting fungus[J]. Current Science, 2000, 79(11):1548-1554. https://www.researchgate.net/publication/202000840_Plant_productivity_determinants_beyond_minerals_water_and_light_Piriformospora_indica_A_revolutionary_plant_promoting_fungus [7] LEE Y C, JOHNSON J M, CHIEN C T, et al. Growth promotion of Chinese cabbage and Arabidopsis by Piriformospora indica is not stimulated by mycelium-synthesized auxin[J]. Molecular Plant-Microbe Interactions, 2011, 24(4):421-431. doi: 10.1094/MPMI-05-10-0110 [8] SIRRENBERG A, GÖBEL C, GROND S, et al. Pirifor-mospora indica affects plant growth by auxin production[J]. Physiol Plant, 2007, 131(4):581-589. doi: 10.1111/ppl.2007.131.issue-4 [9] 陈佑源, 楼兵干, 高其康, 等.印度梨形孢诱导油菜抗旱性机理的初步研究[J].农业生物技术学报, 2013, 21(3):272-281. doi: 10.3969/j.issn.1674-7968.2013.03.003CHEN Y Y, LOU B G, GAO Q K, et al. Preliminary study on mechanisms of drought resistance in Brassica napus L. conferred by Piriformospora indica[J]. Journal of Agricultural Biotechnology, 2013, 21(3):272-281.(in Chinese) doi: 10.3969/j.issn.1674-7968.2013.03.003 [10] SUN C, JOHNSON J M, CAI D, et al. Piriformospora indica, confers drought tolerance in Chinese cabbage leaves by stimulating antioxidant enzymes, the expression of drought-related genes and the plastid-localized CAS protein[J]. Journal of Plant Physiology, 2010, 167(12):1009-1017. doi: 10.1016/j.jplph.2010.02.013 [11] 张文英, 汪嫒嫒, 蒿若超, 等.印度梨形孢真菌促进芝麻生长并提高芝麻抗旱性[J].中国油料作物学报, 2014, 36(1):71-75, 83. http://d.old.wanfangdata.com.cn/Periodical/zgylzwxb201401011ZHANG W Y, WANG A A, HAO R C, et al. Endophytic fungus Piriformospora indica promotes growth and confers drought tolerance in sesame (Sesamum indicum L.)[J]. Chinese Journal of Oil Crop Sciences, 2014, 36(1):71-75, 83.(in Chinese) http://d.old.wanfangdata.com.cn/Periodical/zgylzwxb201401011 [12] BALTRUSCHAT H, FODOR J, HARRACH B D, et al. Salt tolerance of barley induced by the root endophyte Piriformospora indica is associated with a strong increase in antioxidants[J]. New Phytologist, 2008, 180(2):501-510. doi: 10.1111/nph.2008.180.issue-2 [13] FAKHRO A, ANDRADE-LINARES D R, BARGEN S V, et al. Impact of Piriformospora indica, on tomato growth and on interaction with fungal and viral pathogens[J]. Mycorrhiza, 2010, 20(3):191-200. doi: 10.1007/s00572-009-0279-5 [14] KUMAR M, YADAV V, TUTEJA N, et al. Antioxidant enzyme activities in maize plants colonized with Piriformospora indica[J]. Microbiology, 2009, 155:780-790. doi: 10.1099/mic.0.019869-0 [15] SAHAY N S, VARMA A. Piriformospora indica:a new biological hardening tool for micropropagated plants[J]. Fems Microbiology Letters, 1999, 181(2):297-302. doi: 10.1111/fml.1999.181.issue-2 [16] SAHAY N S, VARMA A. A biological approach towards increasing the rates of survival of micro propagated plants[J]. Current Science, 2000, 78(2):126-129. [17] RAI M, ACHARYA D, SINGH A, et al. Positive growth responses of the medicinal plants Spilanthes calva and Withania somnifera to inoculation by Piriformospora indica in a field trial[J]. Mycorrhiza, 2001, 11(3):123-128. doi: 10.1007/s005720100115 [18] PRASAD R, BAGDE U S, PUSPANGADAN P, et al. Bacopa monniera L.:pharmacological aspects and case study involving Piriformospora indica[J]. International Journal of Integrative Biology, 2008, 3(2):100-110. https://www.researchgate.net/publication/228621233_Bacopa_monniera_L_Pharmacological_Aspects_and_Case_Study_Involving_Piriformospora_indica [19] RAI M, VARMA A. Arbuscular mycorrhiza-like biotechnological potential of Piriformospora indica, which promotes the growth of Adhatoda vasica Nees[J]. Electronic Journal of Biotechnology, 2008, 8(1):107-112. http://www.wanfangdata.com.cn/details/detail.do?_type=perio&id=Open J-Gate000000281989 [20] BALDI A, JAIN A, GUPTA N, et al. Co-culture of arbuscular mycorrhiza-like fungi (Piriformospora indica, and Sebacina vermifera) with plant cells of Linum album, for enhanced production of podophyllotoxins:a first report[J]. Biotechnology Letters, 2008, 30(9):1671-1677. doi: 10.1007/s10529-008-9736-z [21] KÄFER E. Meiotic and mitotic recombination in Aspergillus, and its chromosomal aberrations[J]. Advances in Genetics, 1977, 19:33-131. doi: 10.1016/S0065-2660(08)60245-X [22] MURASHIGE T, SKOOG F. A revised medium for rapid growth and bio assays with tobacco tissue cultures[J]. Physiologia Plantarum, 1962, 15(3):473-497. doi: 10.1111/ppl.1962.15.issue-3 [23] 孙盈盈.测量培养细胞鲜重和干重的新方法[J].生物技术通报, 1997(2):33-34. http://www.wanfangdata.com.cn/details/detail.do?_type=perio&id=QK199700513153SUN Y Y. A new method for measuring fresh weight and dry weight of cultured cells[J]. Biotechnology Bulletin, 1997(2):33-34.(in Chinese) http://www.wanfangdata.com.cn/details/detail.do?_type=perio&id=QK199700513153 [24] 洪法水, 魏正贵, 赵贵文.菠菜叶绿素的浸提和协同萃取反应[J].应用化学, 2001, 18(7):532-535. doi: 10.3969/j.issn.1000-0518.2001.07.007Hong F S, WEI Z G, ZHAO G W. Soaking and synergistic extracting reaction of chlorophyll from spinach[J]. Chinese Journal of Applied Chemistry, 2001, 18(7):532-535.(in Chinese) doi: 10.3969/j.issn.1000-0518.2001.07.007 [25] PHAM G H. Axenic culture of symbiotic fungus Piriformospora indica[C]//VARMA A. Plant Surface Microbiology. Berlin: Springer-Verlag, 2004: 593-613. https://www.researchgate.net/publication/283384392_Axenic_Culture_of_Symbiotic_Fungus_Piriformospora_indica -
点击查看大图
计量
- 文章访问数: 2075
- HTML全文浏览量: 460
- PDF下载量: 71
- 被引次数: 0
