Induction Effect of Colchicine on Polyploidy of Medicago polymorpha

doi:10.11924/j.issn.1000-6850.casb2025-0045

Abstract

Abstract:

Using the seeds and plantlets of Medicago polymorpha in Shanghai as experimental materials, different concentrations of colchicine were set (0%, 0.01%, 0.03%, 0.05%, 0.07%, 0.09%, 0.11%), and plant tissue culture, plant biochemical physiology, and plant molecular biology techniques were used to investigate the effects of colchicine treatment on the germination of seeds, the growth and development of plantlets, as well as related physiological and biochemical indicators and gene expression in M. polymorpha. The results showed that the addition of colchicine and colchicine soaking to MS medium could inhibit the germination rate of M. polymorpha seeds and the plant height of germinated seedlings, reduce their leaf number, increase their stem thickness, promote their rooting, increase their CAT activity, SOD activity, POD activity, MDA content, soluble sugar content, soluble protein content, and proline content, promote their upregulation of WRKY transcription factor, protein TIFY 8, transcription factor MYB13, NAC transcription factor 47, ABC transporter, late embryogenesis rich protein 2, and ethylene responsive transcription factor ERF110 gene expression. The addition of colchicine to MS medium and soaking in colchicine could inhibit the plant height, leaf number, stem diameter, root number, and root length of plantlets of M. polymorpha, promote their increase of stem diameter, CAT activity, SOD activity, POD activity, MDA content, soluble sugar content, soluble protein content, and proline content of test tube seedlings of M. polymorpha, and promote their upregulation of gene expression of WRKY transcription factor, protein TIFY 8, transcription factor MYB13, NAC transcription factor 47, ABC transporter, late embryogenesis enriched protein 2, and ethylene responsive transcription factor ERF110. The polyploidy induction effect of M. polymorpha seeds on colchicine added medium was lower than the polyploidy induction effect of M. polymorpha seeds soaked in colchicine, the polyploidy induction effect of M. polymorpha seeds soaked in colchicine was lower than the polyploidy induction effect of M. polymorpha plantlets on colchicine added medium, the polyploidy induction effect of M. polymorpha plantlets on colchicine added medium was lower than the polyploidy induction effect of M. polymorpha plantlets soaked in colchicine. The results of this experiment had practical significance for the induction of polyploidy in M. polymorpha, and could provide theoretical support for the study of signal pathways for polyploidy induction in M. polymorpha and the application of colchicine in inducing polyploidy in M. polymorpha.

Key words: colchicine, Medicago polymorpha, seed germination, plantlet, physiological and biochemical indexes, gene expression

HONG Senrong, JI Lili. Induction Effect of Colchicine on Polyploidy of Medicago polymorpha[J]. Chinese Agricultural Science Bulletin, 2025, 41(22): 13-26.

Figures/Tables 18

References 42

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秋水仙素浓度/%	萌发率/%	成苗率/%	黄化率/%	根数/条	根长/cm
CK	96.30±3.71Aa	40.12±1.53Aa	5.240±1.673Bb	8.33±1.04Aa	0.69±0.05Aa
0.01	98.81±2.06Aa	47.29±2.89Aa	6.680±1.682Bb	0.00±0.00Bb	0.00±0.00Bb
0.03	94.08±5.65Aa	21.27±3.62Bb	11.49±2.774Aa	0.00±0.00Bb	0.00±0.00Bb
0.05	93.80±10.43Aa	25.37±1.30Bb	17.36±1.362Aa	0.00±0.00Bb	0.00±0.00Bb
0.07	91.68±8.02Aa	28.97±2.50Bb	14.68±1.741Aa	0.00±0.00Bb	0.00±0.00Bb
0.09	88.83±3.05Aab	21.90±2.79Bb	18.41±2.784Aa	0.00±0.00Bb	0.00±0.00Bb
0.11	86.29±3.92Abc	22.15±4.11Bb	14.56±1.896Aa	0.00±0.00Bb	0.00±0.00Bb

秋水仙素浓度/%	萌发率/%	成苗率/%	黄化率/%	根数/条	根长/cm
CK	96.30±3.71Aa	40.12±1.53Aa	5.240±1.673Bb	8.33±1.04Aa	0.69±0.05Aa
0.01	98.81±2.06Aa	47.29±2.89Aa	6.680±1.682Bb	0.00±0.00Bb	0.00±0.00Bb
0.03	94.08±5.65Aa	21.27±3.62Bb	11.49±2.774Aa	0.00±0.00Bb	0.00±0.00Bb
0.05	93.80±10.43Aa	25.37±1.30Bb	17.36±1.362Aa	0.00±0.00Bb	0.00±0.00Bb
0.07	91.68±8.02Aa	28.97±2.50Bb	14.68±1.741Aa	0.00±0.00Bb	0.00±0.00Bb
0.09	88.83±3.05Aab	21.90±2.79Bb	18.41±2.784Aa	0.00±0.00Bb	0.00±0.00Bb
0.11	86.29±3.92Abc	22.15±4.11Bb	14.56±1.896Aa	0.00±0.00Bb	0.00±0.00Bb

秋水仙素浓度/%	株高/cm	叶片数/片	茎粗/cm	根数/条	根长/cm
CK	6.00±0.12Aa	11.67±1.66Aa	0.10±0.01Cc	8.67±0.67Aa	3.13±0.27Aa
0.01	0.93±0.03Bb	10.33±0.15Aa	0.12±0.01BCc	0.00±0.00Ee	0.00±0.00Cc
0.03	1.02±0.06Bb	6.50±0.50Bb	0.16±0.02ABb	1.00±0.13Bb	0.23±0.11Bb
0.05	0.97±0.05Bb	2.08±0.13Cc	0.28±0.01Aa	0.66±0.08Cc	0.33±0.12Bb
0.07	0.83±0.03Bb	2.12±0.34Cc	0.26±0.04Aa	0.33±0.02Dd	0.20±0.01Bb
0.09	0.80±0.08Bb	2.24±0.31Cc	0.26±0.05Aa	0.32±0.03Dd	0.31±0.04Bb
0.11	0.80±0.03Bb	2.31±0.24Cc	0.23±0.03Aa	0.33±0.01Dd	0.36±0.02Bb

秋水仙素浓度/%	株高/cm	叶片数/片	茎粗/cm	根数/条	根长/cm
CK	6.00±0.12Aa	11.67±1.66Aa	0.10±0.01Cc	8.67±0.67Aa	3.13±0.27Aa
0.01	0.93±0.03Bb	10.33±0.15Aa	0.12±0.01BCc	0.00±0.00Ee	0.00±0.00Cc
0.03	1.02±0.06Bb	6.50±0.50Bb	0.16±0.02ABb	1.00±0.13Bb	0.23±0.11Bb
0.05	0.97±0.05Bb	2.08±0.13Cc	0.28±0.01Aa	0.66±0.08Cc	0.33±0.12Bb
0.07	0.83±0.03Bb	2.12±0.34Cc	0.26±0.04Aa	0.33±0.02Dd	0.20±0.01Bb
0.09	0.80±0.08Bb	2.24±0.31Cc	0.26±0.05Aa	0.32±0.03Dd	0.31±0.04Bb
0.11	0.80±0.03Bb	2.31±0.24Cc	0.23±0.03Aa	0.33±0.01Dd	0.36±0.02Bb

秋水仙素浓度/ %	CAT活性/ [U/(min·g·FW)]	SOD活性/ [U/(min·g·FW)]	POD活性/ [U/(min·g·FW)]	MDA含量/ [µmol/(g·FW)]	可溶性糖含量/ [mg/(g·FW)]	可溶性蛋白含量/ [mg/(g·FW)]	脯氨酸含量/ [µg/(g·FW)]
CK	169.35±25.62Ff	238.12±48.21Ee	856.39±56.32Ee	5.59±0.69Cc	3.46±0.63Dd	26.39±2.36Ee	125.49±23.61Gg
0.01	234.96±54.21Dd	364.95±26.31Cc	981.06±52.42Dd	5.36±0.86Cc	5.96±0.85Cc	36.29±5.12Dd	234.81±52.13Ff
0.03	316.95±41.87Bb	425.96±28.91Bb	1325.42±48.75Cc	12.63±1.36Bb	8.65±1.26Bb	42.55±1.85Cc	352.64±49.87Cc
0.05	423.82±28.19Aa	498.67±47.26Aa	1624.82±62.13Aa	24.19±3.24Aa	12.84±1.64Aa	52.61±4.23Aa	428.51±51.22Aa
0.07	263.48±22.11Cc	413.54±31.54Bb	1482.44±82.13Bb	22.43±3.16Aa	13.96±2.85Aa	46.82±5.12Bb	382.19±61.23Bb
0.09	244.85±32.16Dd	342.68±24.16Cc	1023.52±77.39Dd	25.21±1.96Aa	7.86±1.03Bb	45.62±1.69Bb	316.74±31.85Dd
0.11	219.77±35.49Ee	318.49±19.85Dd	989.67±49.88Dd	26.78±2.88Aa	7.94±0.96Bb	41.84±2.51Cc	271.99±27.49Ee