Effects of exogenous BR and IAA on drought tolerance of Populus deltoides × P.nigra
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摘要: 为了探究外源油菜素类固醇(brassinosteroid，BR)和IAA对杨树耐旱性的影响，本文选取3种欧美杨无性系NE19、R270、107的1年生盆栽苗木，分别喷洒10 μmol/L BR和100 mg/L IAA，并同时控制土壤含水量为20%，对其进行为期15 d的干旱处理和非干旱处理，对它们的生长和生理变化进行检测。结果表明：与无激素组相比，BR与IAA均可显著提高干旱条件下3种欧美杨无性系的净光合速率(Pn)、光化学量子产量(Fv/Fm)、叶绿素含量、叶片保水能力。BR处理NE19、R270、107的Pn比非干旱组分别提高19%、60%、85%，IAA处理分别高了35%、50%、80%；对Fv/Fm，BR处理的NE19、R270、107比对照组提高了7.7%、7.5%、10.9%，而IAA组提高了7.4%、8%、11.9%。以上结果说明外源施加BR与IAA提高了欧美杨无性系的耐旱性。统计经过激素处理后的欧美杨无性系气孔指数发现，BR处理NE19、R270、107的气孔指数分别下降12.28%、25.60%、20.31%，IAA处理分别下降13.00%、13.15%、14.48%，因此我们推测施加BR和IAA，会抑制植物气孔发育以降低干旱条件下的水分消耗；通过荧光定量PCR，检测与气孔发育6个相关基因EPF1、EPF2、EPFL9、ERECTA、FAMA、SDD1的表达量特征，发现BR和IAA处理导致负调节气孔发育相关基因EPF1、EPF2、ERECTA、SDD1表达量升高，正调节气孔发育基因FAMA与EPFL9表达量下降。综上所述，植物激素BR与IAA参与欧美杨气孔发育过程，在干旱条件下通过抑制气孔发育提高欧美杨抗旱性。Abstract: In order to study the effects of exogenous BR and IAA on the drought resistance of poplar, the 1-year-old potted seedlings of three Populus deltoides × P. nigra clones, NE19, R270, and 107 were selected for 15 days of treatment and sprayed with 10 μmol/L brassinostevoid (BR) and 100 mg/L IAA. The results showed that BR and IAA significantly improved the net photosynthetic rate (Pn), photochemical quantum yield (Fv/Fm), chlorophyll content, leaf water-holding capacity under drought condition compared with the hormone-free. The Pn of NE19, R270 and 107 was increased by 19%, 60% and 85%, respectively in BR treatment, and 35%, 50% and 80%, respectively in IAA treatment. The Fv/Fm of BR group increased by 7.7%, 7.5% and 10.9% compared with the non-hormone group, while the IAA group increased by 7.4%, 8% and 11.9%, respectively. The results indicated that exogenous BR and IAA improved the drought tolerance of Populus deltoides × P. nigra clones. Examining the leaf discs treated with exogenous hormones treatment and it was indicated that, for all three clones, the stomatal indix decreased by 12.28%, 25.60% and 20.31%, respectively in BR treatment, and the IAA treatment decreased by 13.00%, 13.15% and 14.48%, respectively. BR and IAA treatments inhibit stomatal development to reduce water loss under drought conditions.The expressions of EPF1, EPF2, EPFL9, ERECTA, FAMA, SDD1 in stomatal growth-related genes were detected by fluorescence quantitative PCR. EPF1, EPF2, ERECTA and SDD1 which negatively control stomatal development were increased, but the expressions of FAMA and EPFL9 which positively regulate the process were decreased. In summary, phytohormone BR and IAA are involved in the process of stomatal development in Populus deltoides × P.nigra, and increase drought resistance of poplar by inhibiting stomatal development under drought conditions.
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Keywords:
- BR /
- IAA /
- Populus deltoides × P.nigra /
- exogenous hormone /
- drought tolerance
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图 1 不同水分处理条件下3个无性系的光合速率
A、B.土壤相对含水量50%; C、D.土壤相对含水量20%。图中数据为平均值±标准误差(n=3)。下同。
Figure 1. Variation of photosynthetic rate of 3 clones under different water treatments
A, B, soil relative water content is 50%; C, D, soil relative water content is 20%; data are mean ± SD (n=3). NE-19, Populus nigra×(P. deltoides×P. nigra); R270, P. deltoides ×P. nigra, 107, P.×euramericana cv.'74/76'. The same below.
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图 2 不同水分处理条件下外源IAA和BR对3种欧美杨光化学量子产量(Fv/Fm)的影响
Figure 2. Effects of exogenous IAA and BR on the photochemical quantum yield (Fv/Fm) of three clones under different water treatments
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图 3 不同水分处理条件下外源IAA和BR对3个无性系欧美杨叶绿素含量的影响
A~C.3种无性系叶绿素a含量；D~F.3种无性系叶绿素b含量。
Figure 3. Effects of exogenous IAA and BR on chlorophyll content of three clones under different water treatments
A-C, chlorophyll a content of three clones; D-F, chlorophyll b content of three clones.
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图 4 外源施加IAA与BR对欧美杨叶片相对含水量的影响
Figure 4. Effects of exogenous IAA and BR on leaf relative water content of Populus deltoides × P. nigra
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图 5 外源施加IAA与BR对3个欧美杨无性系气孔导度(Gs)的影响
Figure 5. Effects of exogenous IAA and BR onstomatal conductance of three clones
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图 6 IAA与BR降低欧美杨幼叶气孔密度
Figure 6. IAA and BR reducing the stomatal density of Populus deltoides × P. nigra
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图 7 IAA与BR处理下气孔发育相关基因表达变化
Figure 7. Transcriptional abundance of different stomatal development-related genes compared with hormone-free group
表 1 各处理组详情表
Table 1 Treatment group in detail
土壤相对含水量
Soil relative water content20% 50% NE19 Hormone free-NE19 Hormone free-NE19 IAA-NE19 IAA-NE19 BR-NE19 BR-NE19 R270 Hormone free-R270 Hormone free-R270 IAA- R270 IAA- R270 BR- R270 BR- R270 107 Hormone free-107 Hormone free-107 IAA-107 IAA-107 BR-107 BR-107 
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表 2 荧光定量PCR引物
Table 2 Primers for quantitative real-time RT-PCR
基因名称Gene name 正向引物Forward primer 反向引物Reverse primer UBQ AGACCTACACCAAGCCCAAGAAGAT CCAGCACCGCACTCAGCATTAG Potri.019G102800 ATGCTCACCATGTAGACTAG GGACAGGATAAGACTTGTTATG Potri.013G116600 GGAAGCAGCAAGAGAAGA TGAAGGCACATGGTAGTATT Potri.002G249900 CACCACCTGCTCATAAGTT GACATCTGCTCTTCATTGC Potri.013g136100 TCTCTGTCTTCTTCCAATGG AGTCTCAGCCTCTTCAAGT ERECTA ATCCAGGGCTGATGACAACA ACAGTAATGCAAGTTGGAAA FAMA ATCAGTGCCAAGCTTGAAGA AACACAGGGCAGTTGCTTCC SDD1 AATATCATGTCAGGATCATC TTGTCACTAACATAGGCAGT
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表 3 3个无性系各处理间气孔指数对比
Table 3 Variation of stomatal index among 3 clone treatments
无激素
Hormone-freeIAA BR IAA组降低率
Reducing rate of IAA treatment/%BR组降低率
Reducing rate of BR treatment/%NE19 16.61+0.85a 14.45+0.45b 14.57+0.96b 13.00 12.28 R270 20.00+0.99a 17.37+0.22b 14.88+0.7c 13.15 25.60 107 18.51+1.3a 15.83+0.71b 14.75+0.89b 14.48 20.31 注：表中数据为平均值±标准误差(n=3)，不同小写字母代表差异显著(P＜0.05)。Notes: data are mean ± SD (n=3).Different letters indicate significant difference (P＜0.05).
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