BIOLOGICAL ACTIVITY AND QUANTIFICATION OF POTENTIAL AUTOTOXINS FROM THE LEAVES OF PICEA SCHRENKIANA

doi:10.3773/j.issn.1005-264x.2009.01.021

Abstract

Abstract:

Aims Schrenk spruce (Picea schrenkiana) forest is an important zonal vegetation in Tianshan Mountain, northwestern China. In addition to being an important timber resource, P. schrenkiana has important soil and water conservation functions and makes up 98.6% of the total stand volume in the region. Schrenk spruce forest exhibits slow growth, regeneration obstacles, declining productivity and confusing size and age structure. Our aims were to 1) compare autotoxicities of three organic fractions from leaves and 2) identify autotoxins from leaves. Findings could improve understanding the mechanisms of P. schrenkiana autotoxicity.
Methods Fresh leaves were extracted with water and partitioned against diethyl ether, ethyl acetate and n-butanol sequentially. We tested the three organic fractions for inhibitory activity, using germination rate, vigor and index as germination parameters and radicle and plumule elongation and weight (wet and dry) change of seedlings as seedling growth parameters. We investigated the chemical nature of the three fractions using GC-MS-MS and NMR. The chemicals in the samples were estimated on the basis of ion-molecular messenger of MS-MS spectral of samples and MS-MS user library spectra of pure reference compounds. These findings were confirmed by comparing the retention time and mass spectral in the GC-MS-MS of samples with those of reference standards under the same conditions.
Important findings The diethyl ether fraction exhibited the strongest phytotoxicity, reducing seed germination at low concentration (IC₅₀ = 5.84 mg·ml^-1), and n-butanol had the weakest effect (IC₅₀ > 10.00 mg·ml^-1). The diethyl ether fraction had a significant inhibitory effect on seedling growth at a concentration of 1.25 mg·ml^-1, whereas, the n-butanol fraction at 1.25 mg·ml^-1 stimulated it and at 2.5 mg·ml^-1 inhibited it. Seventeen secondary metabolites were identified and quantified in the three fractions, including 2-keto-4a-methyl-8-methoxy-2,3,4,4a,5,6,11,12-ocahydro chrysene in the diethyl ether fraction (this metabolite had been found but not previously reported as an allelochemical). When we used identified components to prepare synthetic mixtures, it showed the same inhibitory effect on seed germination and seedling growth as did the three fractions.

Key words: autotoxicity, Picea schrenkiana, extracts, germination index

PAN Cun-De, WANG Qiang, RUAN Xiao, LI Zhao-Hui. BIOLOGICAL ACTIVITY AND QUANTIFICATION OF POTENTIAL AUTOTOXINS FROM THE LEAVES OF PICEA SCHRENKIANA[J]. Chin J Plant Ecol, 2009, 33(1): 186-196.

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Figures/Tables 10

References 31

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	浓度Concentrations (mg·ml^-1)	发芽率 Germination rate ( % )	发芽势 Germination vigor ( % )	发芽指数 Germination index
乙醚萃取物 Diethyl ether extract	1.25	74.37±6.43^B	57.73±5.37^C	30.35±1.71^C
	2.50	64.60±3.86^CD	45.87±5.40^DE	23.46±1.53^DE
	5.00	47.90±2.10^F	25.10±4.35^F	15.31±2.15^F
	10.00	26.67±6.86^H	12.17±4.05^G	5.38±0.83^H
乙酸乙酯萃取物 Ethyl acetate extracts	1.25	84.13±2.21^A	66.13±2.02^B	40.65±1.38^B
	2.50	71.93±3.43^B	55.00±2.85^C	29.29±0.88^C
	5.00	54.87±6.69^EF	41.80±5.43^DE	20.92±1.44^E
	10.00	39.40±2.94^G	26.47±3.32^F	12.15±1.24^G
正丁醇萃取物 n-butanol extract	1.25	88.83±3.07^A	74.07±2.06^A	44.79±1.42^A
	2.50	68.03±4.99^BC	55.13±2.78^C	24.86±2.67^D
	5.00	60.43±3.89^DE	47.67±4.93^D	22.95±2.44^DE
	10.00	51.80±6.78^F	39.43±1.72^E	14.83±0.99^FG
CK	CK	86.67±1.15^A	70.67±7.02^AB	43.49±2.13^AB

	浓度Concentrations (mg·ml^-1)	发芽率 Germination rate ( % )	发芽势 Germination vigor ( % )	发芽指数 Germination index
乙醚萃取物 Diethyl ether extract	1.25	74.37±6.43^B	57.73±5.37^C	30.35±1.71^C
	2.50	64.60±3.86^CD	45.87±5.40^DE	23.46±1.53^DE
	5.00	47.90±2.10^F	25.10±4.35^F	15.31±2.15^F
	10.00	26.67±6.86^H	12.17±4.05^G	5.38±0.83^H
乙酸乙酯萃取物 Ethyl acetate extracts	1.25	84.13±2.21^A	66.13±2.02^B	40.65±1.38^B
	2.50	71.93±3.43^B	55.00±2.85^C	29.29±0.88^C
	5.00	54.87±6.69^EF	41.80±5.43^DE	20.92±1.44^E
	10.00	39.40±2.94^G	26.47±3.32^F	12.15±1.24^G
正丁醇萃取物 n-butanol extract	1.25	88.83±3.07^A	74.07±2.06^A	44.79±1.42^A
	2.50	68.03±4.99^BC	55.13±2.78^C	24.86±2.67^D
	5.00	60.43±3.89^DE	47.67±4.93^D	22.95±2.44^DE
	10.00	51.80±6.78^F	39.43±1.72^E	14.83±0.99^FG
CK	CK	86.67±1.15^A	70.67±7.02^AB	43.49±2.13^AB

乙醚萃取物 Diethyl ether extract (mg·ml^-1)	干重 Dry weight (g)	乙酸乙酯萃取物 Ethyl acetate extract (mg·ml^-1)	干重 Dry weight (g)	正丁醇萃取物 n-butanol extract (mg·ml^-1)	干重 Dry weight (g)
1.25	0.041±0.003^A	1.25	0.041±0.003^A	1.25	0.041±0.002^A
2.50	0.041±0.001^A	2.50	0.041±0.002^A	2.50	0.040±0.002^A
5.00	0.042±0.002^A	5.00	0.040±0.002^A	5.00	0.042±0.002^A
10.00	0.041±0.002^A	10.00	0.042±0.001^A	10.00	0.040±0.002^A
CK	0.040±0.001^A	CK	0.040±0.001^A	CK	0.040±0.001^A

乙醚萃取物 Diethyl ether extract (mg·ml^-1)	干重 Dry weight (g)	乙酸乙酯萃取物 Ethyl acetate extract (mg·ml^-1)	干重 Dry weight (g)	正丁醇萃取物 n-butanol extract (mg·ml^-1)	干重 Dry weight (g)
1.25	0.041±0.003^A	1.25	0.041±0.003^A	1.25	0.041±0.002^A
2.50	0.041±0.001^A	2.50	0.041±0.002^A	2.50	0.040±0.002^A
5.00	0.042±0.002^A	5.00	0.040±0.002^A	5.00	0.042±0.002^A
10.00	0.041±0.002^A	10.00	0.042±0.001^A	10.00	0.040±0.002^A
CK	0.040±0.001^A	CK	0.040±0.001^A	CK	0.040±0.001^A

保留时间 Retention time (min)	化合物 Chemical constituents	相对含量Relative amount (%)	化学文摘登记号 CAS	前体物离子 Precursor ion (m/z)	定量物离子 Quantization ion (m/z)
9.76	对羟基苯甲酸p-hydroxybenzoic acid	8.19	99-96-7	267	223
13.53	云杉酮Schrenkianaone	20.32	102323-79-5	294	279
14.53	肉桂酸 t-cinnamic acid	9.98	140-10-3	205	159
15.98	脱落酸 ABA	30.47	14375-45-2	293	249
17.32	槲皮素 Quercetin	14.49	6151-25-3	587	466
—	其他未知成分Other unknown components	16.55