四倍体海稻86的诱导、鉴定及其耐盐碱特性评价分析

doi:10.3724/SP.J.1006.2024.32035

Abstract

Abstract:

Polyploidization is an important trend in plant evolution. Polyploid plants often have stronger resistance to stress than diploid ones. Haidao 86 is a rice germplasm resource with strong salt-alkali tolerance, which has important utilization value for the application of saline-alkaline land and the increase of grain yield. To make full use of the advantages of enhanced stress resistance in polyploid plants and breed new tetraploid rice varieties with stronger salt-alkali tolerance, diploid Haidao 86 was submitted for in vitro chromosome doubling to create autotetraploid Haidao 86 in this study. After stress treatment with NaCl, Na₂CO₃, and PEG-6000, the phenotypic indexes, physiological and biochemical indexes of tetraploid and diploid Haidao86 at germination and seedling stages were detected to understand the salt-alkali tolerance characteristics and differences between them. The results showed as follows: (1) By in vitro chromosome doubling, tetraploid plants can be efficiently induced, with a doubling rate of 27.63%. (2) Compared with diploid Haidao 86, the nuclear DNA content and root tip chromosome number of tetraploid Haidao 86 was doubled. There were significant changes in morphology and agronomic traits, such as plants becoming shorter, stems becoming thicker, effective panicles per plant decreasing, grain and thousand grain weight increasing, total grains per panicle, and filled grains per panicle decreasing, and seed setting rate decreasing. (3) At germination stage, the germination energy, germination rate, shoot length, root length, root number, and water content of tetraploid Haidao 86 were the highest, followed by diploid Haidao 86, and the control Huanghuazhan was the lowest. Tetraploid Haidao 86 had the lowest salt alkali damage rate and grade, followed by diploid Haidao 86, and the control Huanghuazhan had the highest salt alkali damage rate and grade. (4) At seedling stage, tetraploid Haidao 86 had the highest proline and chlorophyll content, the lowest malondialdehyde content and relative electrical conductivity, the highest superoxide dismutase and peroxidase activity, followed by diploid Haidao 86, and the control Huanghuazhan had the lowest proline and chlorophyll content, the highest malondialdehyde content and relative electrical conductivity, and the lowest superoxide dismutase and peroxidase activity. So tetraploid Haidao 86 had significant advantages in salt-alkali tolerance in both phenotypic indexes, and physiological and biochemical indexes, and had stronger salt-alkali tolerance than diploid Haidao 86. The results lay the foundation for in-depth research on the salt-alkali tolerance mechanism of Haidao 86, and provide the material and theoretical basis for the breeding of new salt-alkali resistant tetraploid rice varieties.

Key words: Haidao 86, in vitro chromosome doubling, tetraploid, salt-alkali tolerance, germination energy, germination rate, physiological and biochemical indexes

LI Hang-Yu, LIU Xin-Cheng, HE Wen-Ting, LIU Ke-Yi, QIAO Zhen-Hua, LYU Pin-Cang, ZHANG Xian-Hua, HE Yu-Chi, CAI De-Tian, SONG Zhao-Jian. Induction, identification and salt-alkali tolerance evaluation of tetraploid Haidao 86[J].Acta Agronomica Sinica, 2024, 50(4): 914-931.

Figures/Tables 11

Table 1

Fig. 1

Fig. 2

Table 2

Fig. 3

Fig. 4

Table 3

Table 4

Relative shoot length, root length and root number under saline, alkaline and osmotic stress (%)"

胁迫物质及浓度Stress substance and concentration (mmol L^-1)	相对芽长Relative shoot length			相对根长Relative root length			相对根数Relative root number
胁迫物质及浓度Stress substance and concentration (mmol L^-1)	HD86-2x	HD86-4x	HHZ	HD86-2x	HD86-4x	HHZ	HD86-2x	HD86-4x	HHZ
NaCl 50	86.86±3.00 a	93.75±6.10 a	59.60±12.04 b	76.58±5.58 a	96.65±14.38 a	51.23±13.49 b	89.17±13.46 a	89.44±11.11 a	56.71±3.93 b
NaCl 100	81.01±4.86 a b	88.98±11.51 a	50.92±23.88 b	80.30±2.22 a	93.28±9.37 a	36.86±12.37 b	81.22±10.28 a	87.10±12.67 a	52.46±0.98 b
NaCl 150	83.24±20.63 a	74.64±4.74 a	25.35±9.40 b	62.80±3.28 a	89.83±22.34 a	32.45±5.34 b	78.25±6.34 a	66.51±8.85 a	37.84±1.85 b
NaCl 200	49.98±9.74 ab	61.45±12.82 a	32.58±12.31 b	48.64±4.19 b	78.50±6.70 a	21.31±5.91 c	59.33±13.05	57.06±12.83	37.92±2.33
NaCl 250	64.94±9.10 a	57.09±6.16 a	22.22±5.35 b	43.65±9.55 ab	59.70±18.43 a	18.90±4.82 b	51.09±12.33 a	43.03±5.50 ab	21.23±15.89 b
Na₂CO₃ 10	97.12±2.47	96.80±9.56	96.62±5.14	79.52±3.85	95.11±12.89	80.23±21.33	90.42±8.66 a	86.63±5.80 ab	77.05±4.87 b
Na₂CO₃ 20	66.32±14.34 b	88.97±7.24 a	67.70±5.40 b	57.47±9.61	70.37±3.71	60.30±10.03	83.43±19.51 a	87.11±9.09 a	51.48±7.93 b
Na₂CO₃ 30	63.50±9.61 ab	82.51±18.61 a	36.88±11.40 b	42.63±4.53 ab	58.80±15.00 a	35.17±3.55 b	51.81±17.25 a	68.47±10.08 a	25.73±4.09 b
Na₂CO₃ 40	55.97±15.93 a	69.39±21.32 a	16.39±.9.28 b	28.19±8.24	41.77±17.01	21.36±7.12	48.44±8.87 a	39.28±5.38 a	20.64±5.12 b
Na₂CO₃ 50	39.07±5.72 a	29.11±9.60 a	11.03±1.78 b	30.71±2.87 a	36.08±5.34 a	11.67±3.02 b	25.71±4.32	21.22±7.14	16.64±5.09
PEG 10	80.68±8.24 b	99.01±10.10 a	70.82±3.40 b	53.87±2.20 b	87.56±12.83 a	67.79±6.70 b	58.56±11.38 b	81.18±7.26 a	63.69±4.84 b
PEG 20	76.05±2.56 ab	90.24±19.11 a	58.52±4.83 b	37.31±3.74	51.96±15.36	31.34±18.60	46.48±8.52	45.22±3.21	57.01±1.65
PEG 30	30.35±3.86 b	84.38±16.76 a	55.23±13.28 b	30.80±5.95 ab	38.83±4.71 a	19.17±7.68 b	28.56±11.12	35.46±5.00	28.00±3.28
PEG 40	17.46±4.03 b	37.99±3.66 a	22.05±4.50 b	13.98±2.12	21.13±4.22	25.47±12.35	17.01±5.59	20.34±4.12	13.41±2.99
PEG 50	0±0.00 b	24.44±2.88 a	0±0.00 b	0±0.00 b	29.06±12.64 a	0±0.00 b	0±0.00 b	9.73±1.55 a	0±0.00 b

Table 4

Fig. 5

Fig. 6

Fig. 7

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批次 Batch	种胚数 NE	愈伤组织数 NC	诱导率 IR (%)	加倍存活数 NSC	加倍存活率 SR (%)	分化愈伤组织数 NDC	分化率 DR (%)	总成苗数 TSN	四倍体苗数 PSN	加倍率CDR (%)
1	80	70	87.50	70	100	39	55.71	16	5	31.25
2	75	64	85.33	64	100	47	73.44	21	6	28.57
3	75	54	72.00	46	85.19	29	63.04	13	3	23.08
均值 Mean ± SD	76.67 ±2.89	62.67 ±8.08	81.61 ±8.39	60.00 ±12.49	95.06 ±8.55	38.33 ±9.02	64.06 ±8.91	16.67 ±4.04	4.67 ±1.53	27.63 ±4.16

材料 Material	株高 PH (cm)	单株有效穗数 EP	穗长 PL (cm)	粒长 GL (mm)	粒宽 GW (mm)
HD86-2x	181.96±3.34	15.40±1.14	26.79±1.87	7.01±0.04	3.03±0.04
HD86-4x	152.40±4.18^**	10.20±1.30^**	23.36±1.85^*	9.04±0.06^**	4.02±0.03^**
材料 Material	芒长 AL (cm)	每穗总粒数 TG	每穗实粒数 FG	结实率 SSR (%)	千粒重 TGW (g)
HD86-2x	0.10-6.60	162.72±5.96	147.16±7.66	90.41±2.29	24.90±1.05
HD86-4x	0.10-7.20	86.60±9.44^**	28.34±4.25^**	32.74±3.41^**	33.94±1.37^**

胁迫物质及浓度 Stress substance and concentration (mmol L^-1)	HD86-2x		HD86-4x		HHZ
胁迫物质及浓度 Stress substance and concentration (mmol L^-1)	盐碱害率 Damage rate (%)	等级Grade	盐碱害率 Damage rate (%)	等级Grade	盐碱害率 Damage rate (%)	等级Grade
0 (CK)	0±0.00	1	0±0.00	1	0±0.00	1
NaCl 50	0±0.00 b	1	0±0.00 b	1	14.43±1.96 a	1
NaCl 100	1.47±1.68 b	1	1.37±1.72 b	1	17.63±2.06 a	1
NaCl 150	13.97±1.15 a	1	3.17±3.37 b	1	22.07±7.13 a	3
NaCl 200	17.97±1.78 ab	1	9.10±2.10 b	1	29.43±10.86 a	3
NaCl 250	23.70±3.92 b	3	11.73±1.66 b	1	42.60±10.10 a	5
Na₂CO₃ 10	7.80±1.91 b	1	7.80±1.91 b	1	16.67±3.35 a	1
Na₂CO₃ 20	18.90±1.91 ab	1	7.80±1.91 b	1	28.90±15.76 a	3
Na₂CO₃ 30	26.67±8.84 ab	3	15.57±7.68 b	1	38.90±11.69 a	3
Na₂CO₃ 40	35.53±3.87 b	3	26.67±3.35c	3	60.00±3.30 a	5
Na₂CO₃ 50	54.43±1.96 b	5	32.23±3.87 c	3	73.33±3.35 a	7
PEG 10	11.10±1.91 b	1	0±0.00 c	1	20.00±3.30 a	1
PEG 20	37.80±1.91 b	3	21.10±1.91c	3	55.57±5.10 a	5
PEG 30	47.77±7.74 b	5	44.43±9.64 b	5	77.77±3.87 a	7
PEG 40	81.13±7.68 ab	7	72.20±1.91 b	5	88.90±1.91 a	9
PEG 50	100±0.00 a	9	90.43±2.68 b	9	100.00±0.00 a	9

Induction, identification and salt-alkali tolerance evaluation of tetraploid Haidao 86

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