首页 > 分享 > 接种兰科菌根真菌对不同硝铵比条件下大花蕙兰生长、养分吸收及氮代谢的影响

接种兰科菌根真菌对不同硝铵比条件下大花蕙兰生长、养分吸收及氮代谢的影响

花匠小妙招
2025-05-22 20:33

摘要:

目的

探究兰科菌根真菌(orchid mycorrhizal fungi，OMF)对不同形态氮素配比(NO3−/NH4+)条件下大蕙兰花生长及氮代谢的影响。

方法

以大花蕙兰580和OMF兰科丝核菌C2y1为研究对象，进行盆栽试验。设置土壤浇灌和不浇灌C2y1，每个处理下分别设3个NO3−/NH4+供应比例，即8∶1 (T1)、1∶1 (T2) 和1∶8 (T3)。测定了处理后大花蕙兰幼苗生长、生物量和根系长度的增加情况，分析了处理后叶片中硝酸还原酶(NR)和谷氨酰胺合成酶(GS)活性，采用PCR法检测C2y1定植情况。

结果

未接种C2y1条件下，T2处理的大花蕙兰植株鲜重增长量及干物质积累量显著高于T1和T3处理，T3处理的根系增长量显著高于T1和T2处理。PCR分析结果证明，土壤浇灌C2y1菌剂后，C2y1可有效侵入大花蕙兰根系，形成共生关系，且随着铵态氮占比的增加，根系C2y1侵染率显著增加。相比于未接种对照，接种C2y1可显著促进各硝铵比处理大花蕙兰茎叶生长和生物量积累，茎叶的最大提升效果出现在T3处理，而最大根系促生效果出现在T1处理；接种C2y1显著提高了茎叶对大量元素(N、P、K)和微量元素(Fe、Zn、Cu)的吸收，茎叶最高N、P、K和Zn含量出现在T3处理，对根系中各元素含量提升效果较小。接种和未接种C2y1条件下，大花蕙兰幼苗的NR活性都随着硝态氮比例的下降而降低，而GS活性正相反，且接种C2y1后各硝铵比处理幼苗的GS活性均显著高于未接种处理。

结论

大花蕙兰比较喜好硝态氮，但是铵态氮可促进其根系与C2y1形成共生体来适应高铵环境。接种兰科丝核菌C2y1促进了大花蕙兰生长及对营养的吸收和运转，且在高铵环境下的促生效应大于高硝态氮环境下。

Abstract:

Objectives

This study is to explore the impact of orchid mycorrhizal fungi (OMF) inoculation on the growth and nitrogen metabolism of Cymbidium hybridum under various NO3−/ NH4+ ratios.

Methods

Cultivar Cymbidium hybridum ‘580’ and OMF C2y1 were used as test materials in a pot experiment. The C2y1 inoculation was carried out by watering into soils during the seedling stage of Cymbidium hybridum, taking water as control group. Under each group, N was supplied in three different NO3−/NH4+ ratios of 8∶1 (T1), 1∶1 (T2), and 1∶8 (T3). The increased growth and biomass weight of the shoot, and the enhanced length of roots of C. hybridum seedlings were measured, the nutrient contents in both shoot and root, and the activities of nitrate reductase (NR) and glutamine synthetase (GS) in leaves were analyzed. PCR method was used to confirm the infection of C2y1 fungi.

Results

Without inoculation, the significantly higher fresh weight growth and dry matter accumulation of C. hybridum were recorded under T2, while the significantly higher root growth was recorded at T3. The PCR results proved the infection of C2y1 on the root of C. hybridum through the soil watering method, and the infection intensity was enhanced with the increase of ammonia nitrogen supply ratio. Compared to the no inoculation control, C2y1 inoculation significantly enhanced shoot growth and biomass accumulation of C. hybridum under various nitrogen form ratios, with the largest promotion on shoot growth under T3 treatment, and the highest promotion on root was under T1 treatment. C2y1 inoculation significantly enhanced the absorption of N, P, K, Fe, Zn, Cu. Notably, with the highest concentrations of N, P, K, and Zn in shoot under T1 ratio, while their concentration in roots were not affected by C2y1 inoculation. The NR activity in C. hybridum seedlings under C2y1 inoculation was positively correlated with the nitrate nitrogen supply ratios, while GS activity was on the contrary.

Conclusions

C. hybridum prefers nitrate to ammonia, however, ammonia is conducive the infection of C2y1, which help C. hybridum to adapt to the soil environment. Inoculation of C2y1 could stimulate the growth and nutrient absorption, and the promotion effect under high ammonia environment is even more obvious.

图 1 不同处理下大花蕙兰的整体根段真菌侵染图像

注：a、b、c分别为NO3−/NH4+为8∶1、1∶1、1∶8，不接菌处理，d、e、f分别为NO3−/NH4+为8∶1、1∶1、1∶8，接菌处理。兰花菌根利用台盼蓝进行染色，使用ImageJ图像处理软件对菌根化的大花蕙兰根系染色照片进行处理。

Figure 1. Fungal infection images of root segments of C. hybridum under different treatments

Note: a, b and c are treatments without C2y1 inoculation, with the NO3−/NH4+ ratios being 8∶1, 1∶1 and 1∶8, respectively; d, e and f are the roots of C. hybridum inoculated with C2y1, with the NO3−/NH4+ ratio is 8∶1, 1∶1 and 1∶8, respectively. The orchid mycorrhizae are stained using trypan blue, and the stained images of the mycorrhizal roots of Cymbidium hybridum are processed using ImageJ image-processing software.

图 2 不同硝铵配比下接种C2y1对大花蕙兰植株和根系生长的影响

注：CK—未接种真菌；C2y1—接种真菌。柱上不同小写字母表示处理间差异显著(P<0.05)。右上角表示双因素方差分析结果，*—P<0.05，***—P<0.001。

Figure 2. Effects of inoculation of C2y1 on the growth of above ground parts and roots of C. hybridum seedlings under different nitrate to ammonia supply ratios

Note: CK—No inoculation control; C2y1—Inoculation of C2y1. Different lowercase letters above the bars indicate significant difference among treatments (P<0.05). Top right are two-way ANOVA output, *—P<0.05; ***—P<0.001.

图 3 不同硝铵比下接种C2y1对大花蕙兰叶绿素含量影响

注：CK—未接种真菌；C2y1—接种真菌。柱上不同小写字母表示处理间差异显著(P<0.05)。

Figure 3. Effects of inoculation of C2y1 on the leaf SPAD of C. hybridum under different nitrate to ammonia supply ratios

Note: CK—No inoculation control; C2y1—Inoculation of C2y1. Different lowercase letters above the bars indicate significant difference among treatments (P<0.05).

图 4 不同硝铵配比下接种C2y1对大花蕙兰幼苗养分含量的影响

注：CK—未接种真菌；C2y1—接种真菌。图中不同小写字母表示同一部分处理间差异显著(P<0.05)。

Figure 4. Effects of inoculation of C2y1 on the nutrient concentrations of C. hybridum seedlings under different nitrate to ammonia supply ratios

Note: CK—No inoculation control; C2y1—Inoculation of C2y1. Different lowercase letters in the Fig. indicate significant difference in the element concentrations of the same part among treatments (P<0.05).

图 5 不同硝铵配比下接种C2y1对大花蕙兰幼苗叶片硝酸还原酶和谷氨酰胺合成酶活性的影响

注：CK—未接种真菌；C2y1—接种真菌。柱上不同小写字母表示6个处理间差异显著(P<0.05)。

Figure 5. Effects of C2y1 inoculation on the activities of nitrate reductase (NR) and glutamine synthetase (GS) in the leaves of C. hybridum seedlings under different nitrate to ammonia nitrogen ratios

Note: CK—No inoculation control; C2y1—Inoculation of C2y1. Different lowercase letters above the bars indicate significant difference among the 6 treatments (P<0.05).

图 6 巢氏PCR产物电泳图

注：图(a)为用ITS1/ITS4引物扩增菌根的ITS效果，图(b)为各菌根ITS PCR产物用CITS扩增效果。M为标准，A、B、C分别为未接种处理下NO3−/NH4+为8∶1、1∶1和1∶8处理的菌根，D、E、F分别为接种C2y1处理下8∶1、1∶1和1∶8处理的菌根。

Figure 6. Electrophoretic diagram of nested PCR product

Note: Figure (a) shows the ITS amplification effect of mycorrhiza using the ITS1/ITS4 primer, and (b) shows the CITS amplification effect of ITS PCR products of mycorrhiza. M is marker; A, B, and C are the roots of C. hybridum seedlings without C2y1 inoculation under NO3−/NH4+ ratios of 8∶1, 1∶1, and 1∶8, respectively, and D, E, and F are the roots of C. hybridum seedlings with C2y1 inoculation under NO3−/NH4+ ratios of 8 : 1, 1∶1, and 1∶8, respectively.

表 1 改良的霍格兰营养液配方

Table 1 The formula of modified Hoagland nutrient solution

大量元素
Macro-element化学式
Chemical formula浓度 (mg/L)
Concentration微量元素
Micro-element化学式
Chemical formula浓度 (mg/L)
Concentration PNaH2PO4150.97BH3BO30.56KK2SO4111.54MnMnSO4·H2O0.77CaCaCl269.38ZnZnSO4·7H2O0.22MgMgSO4·7H2O128.38MoNa2MoO4·2H2O0.13CuCuSO4·5H2O0.04FeFeSO4·7H2O4.97

表 2 不同硝铵比下接种C2y1对大花蕙兰生物量及根系菌根侵染率的影响

Table 2 Effects of C2y1 inoculation on the biomass and root fungal infection rate of C. hybridum seedlings under different nitrate to ammonia supply ratios

处理 Treatment生物量 Biomass (g/plant)根系侵染强度 (%)
Infection rate 硝铵比
NO3−/NH4+接种
Inoculation鲜重增长量
Fresh weight increase茎叶干重
Shoot dry weight根干重
Root dry weight 8∶1 (T1)CK1.41±0.01 d0.15±0.00 d0.09±0.00 dnd C2y12.49±0.01 a0.17±0.00 b0.11±0.00 a36.79 1∶1 (T2)CK1.93±0.01 c0.15±0.00 d0.08±0.00 end C2y12.24±0.03 b0.16±0.00 c0.10±0.00 c61.27 1∶8 (T3)CK1.26±0.01 e0.13±0.00 e0.08±0.00 end C2y12.22±0.00 b0.18±0.00 a0.10±0.00 b64.08 显著性 Significance C2y1********* N***ns*** C2y1×N******* 注：CK—未接种真菌；C2y1—接种真菌。数据代表平均值±标准误 (n=30)，同一列数值后不同小写字母表示处理间差异显著 (P<0.05)。nd表示未检测到。*—P<0.05；***—P<0.001；ns—不显著。
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