Burkholderia strains enhance the tolerance of sugarcane to aluminum stress by improving the physiological adaptability and regulating the expression of aluminum responsive genes

doi:10.17521/cjpe.2024.0014

Abstract

Abstract:

Aims Many factors can influence the growth of plants and their adaptability to the growing environment, among which plant growth promoting rhizobacteria (PGPR) may play a key role. Thus, the aim of this study is to explore the rhizosphere microbial resources with high aluminum tolerance and investigate their role in alleviating the aluminum toxicity to sugarcane (Saccharum officinarum).
Methods We isolated and screened the dominant strains of aluminum-tolerant PGPR from the rhizosphere soil of sugarcane and studied their effects on sugarcane growth. By measuring the aluminum accumulation, physiological metabolism and expression changes of aluminum-tolerant genes in the aboveground and underground parts of sugarcane under aluminum stress, we compared the aluminum tolerance of sugarcane inoculated with aluminum-tolerant bacteria and those without aluminum-tolerant bacteria.
Important findings Three strains of dominant sugarcane rhizosphere bacteria, Burkholderia A1, A23 and X6, have strong acid tolerance (pH 3.8) and aluminum tolerance (4 or 5 mmol·L^-1). All the three strains have the ability to dissolve phosphorus, secrete exopolysaccharides (EPS), fix nitrogen and remove Al³⁺. X6 can produce low level of indole-3-acetic acid (IAA), A1 and X6 can secrete a certain amount of iron carriers, and A23 and X6 have 1-aminocyclopropane-l-carboxylic acid (ACC) deaminase activity. The combined bacteria of A1, A23 and X6 improved the plant height, leaf area and fresh mass of aboveground part of sugarcane. Although the bacteria combinations showed no obvious effect on the contents of nitrogen, phosphorus and potassium in aboveground and underground parts of plant, but significantly increased the aluminum content in aboveground part of sugarcane under 0.5 and 1 mmol·L^-1 aluminum stress and significantly reduced the aluminum content in underground part of sugarcane under 1 mmol·L^-1 aluminum stress, and the overall performance was reducing the total aluminum content in sugarcane. The inoculation treatment showed no significant effect on the activities of superoxide dismutase (SOD) and peroxidase (POD), and the contents of proline (Pro) and soluble protein (SP), but significantly reduced the malondialdehyde (MDA) content and significantly increased the catalase (CAT) activity in root tips under aluminum stress, and significantly increased the expression of MAPK and GST genes in root tips and PEPC gene in leaves of sugarcane. These results showed that Al-tolerant Burkholderia strains had significant effects on sugarcane growth, aluminum absorption and accumulation, plant antioxidant defense ability and aluminum-tolerant gene expression, which together improved the aluminum tolerance of sugarcane.

Key words: Burkholderia, Saccharum officinarum, aluminum stress, physiological adaptation, aluminum tolerant gene

LU Zhen, XIE Guang-Jie, Qaisar KHAN, QIN Ying, HUANG Yu-Yan, GUO Dao-Jun, YANG Ting-Ting, YANG Li-Tao, XING Yong-Xiu, LI Yang-Rui, WANG Zhen. Burkholderia strains enhance the tolerance of sugarcane to aluminum stress by improving the physiological adaptability and regulating the expression of aluminum responsive genes[J]. Chin J Plant Ecol, 2025, 49(3): 475-487.

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URL: https://www.plant-ecology.com/EN/10.17521/cjpe.2024.0014

https://www.plant-ecology.com/EN/Y2025/V49/I3/475

Figures/Tables 11

Table 1 Information of primers for quantitative real-time PCR of aluminum response related genes in sugarcane

基因 Gene	上游引物 Forward primer (5′-3′)	下游引物 Reverse primer (5′-3′)
GAPDH	CACGGCCACTGGAAGCA	TCCTCAGGGTTCCTGATGCC
MAPK	CATCGAGCGGATGCTAACCT	AGTCGAACGAGAATGGCTCC
GST	TACATGGCACACCCTGCATT	AGGAACAATACGACGGAGCA
PEPC	TCCACAGGATGAAATGCGCT	TGTAGGGAAGGCGCTGATTG

Table 2 Growth-promoting characteristics of aluminum-tolerant bacteria from the rhizosphere soil of sugarcane (mean ± SD)

菌株 Baterial stain	产IAA能力 Capacity for producing IAA	磷增溶 Phosphorus solubilization (mg·L^-1)	铁载体相对含量 Relative content of siderophore (A/Ar)	固氮 Nitrogen fixation	ACC脱氨酶活性 ACC deaminase activity	胞外多糖 EPS (mg·mL^-1)
A1	-	54.94 ± 2.38^a	0.38 ± 0.03^a	+++	-	0.27 ± 0.03^c
A23	-	59.32 ± 0.99^a	-	+	+	1.29 ± 0.12^b
X6	+	59.42 ± 3.41^a	0.43 ± 0.02^b	+++	+	1.68 ± 0.07^a

Fig. 1 Phylogenetic tree of three strains of aluminum-tolerant bacteria from sugarcane rhizosphere based on 16S rRNA gene.

Fig. 2 Effect of different acidity of culture medium on growth of the aluminum-tolerant bacterial strains (A1, A23, X6) (mean ± SD). Different lowercase letters indicate significant differences (p < 0.05). OD600, the optical density of bacterial suspension at 600 nm.

Fig. 3 Effects of different Al3+ concentrations on the growth of aluminum-tolerant bacterial strains (A1, A23, X6) (mean ± SD). Under the same aluminum concentration treatment, the data on the left represent the OD600 values, and those on the right represent the final pH values. OD600 is the optical density of bacterial suspension at 600 nm, different lowercase letters indicate significant differences (p < 0.05).

Table 3 Changes of residual Al3+ content and pH of culture solution after aluminum-tolerant bacteria from the rhizosphere soil of sugarcane growth 2 days (mean ± SD)

菌株 Bacterial strain	最终pH Final pH	上清液中的Al³⁺浓度 Al³⁺ concentration in supernatant (mmol·L^-1)	剩余Al³⁺百分比 Percentage of residual Al³⁺ (%)
CK	4.50 ± 0.00^c	0.995 ± 0.003^a	99.539 ± 0.003^a
A1	7.91 ± 0.16^a	0.816 ± 0.042^b	81.992 ± 4.038^b
A23	7.02 ± 0.04^b	0.825 ± 0.014^b	82.880 ± 1.331^b
X6	7.79 ± 0.20^a	0.772 ± 0.007^b	77.581 ± 0.830^b

Table 4 Effects of aluminum-tolerant bacterial strain mixture inoculation (T) on agronomic traits of sugarcane (mean ± SD)

处理 Treatment	株高 Plant height (cm)	叶面积 Leaf area (cm²)	干质量 Dry mass (g)		鲜质量 Fresh mass (g)
处理 Treatment	株高 Plant height (cm)	叶面积 Leaf area (cm²)	地上部分 Aboveground part	地下部分 Underground part	地上部分 Aboveground part	地下部分 Underground part
CK	23.53 ± 1.72^b	12.84 ± 1.37^b	5.96 ± 1.14^a	1.06 ± 0.15^a	26.25 ± 6.73^b	8.00 ± 2.83^a
T	29.40 ± 0.47^a	18.14 ± 2.57^a	8.49 ± 1.13^a	1.20 ± 0.09^a	36.80 ± 7.78^a	10.90 ± 0.42^a

Fig. 4 Hematoxylin stained sugarcane root tips. T, aluminum- tolerant bacterial strain mixture inoculation.

Fig. 5 Effects of aluminum-tolerant bacteria inoculation (T) on total nitrogen (A), total phosphorus (B), total potassium (C) and total aluminum (D) contents in shoot and root of sugarcane under aluminum stress (mean ± SD). Different lowercase letters in the same part indicate significant differences among different concentrations and treatments (p < 0.05).

Fig. 6 Effects of aluminum-tolerant bacteria inoculation (T) on superoxide dismutase (SOD) (A), catalase (CAT) (B) and peroxidase (POD) (C) activities and soluble protein (SP) (D), proline (Pro) (E) and malondialdehyde (MDA) (F) contents in sugarcane leaves and root tips under aluminum stress (mean ± SD). Different lowercase letters in the same organ indicate significant differences among different concentrations and treatments (p < 0.05).

Fig. 7 Changes in the expression of aluminum tolerance related genes MAPK (A), GST (B) and PEPC (C) in sugarcane root tips and leaves under aluminum stress (mean ± SD). Different lowercase letters in the same part indicate significant differences among different concentrations and treatments in leaves and roots at the level p < 0.05. T, aluminum-tolerant bacterial strain mixture inoculation.

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