Chinese Bulletin of Botany ›› 2020, Vol. 55 ›› Issue (6): 749-759.DOI: 10.11983/CBB20062
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Yaqin Wang1,2, Ludan Wei1, Wenjing Wang1, Baojun Liu1, Chunling Zhang1, Junwei Zhang1, Yanhong He1,*(
)
HomeReceived:2020-04-16
Accepted:2020-08-26
Online:2020-11-01
Published:2020-11-11
Contact:
Yanhong He
Yaqin Wang, Ludan Wei, Wenjing Wang, Baojun Liu, Chunling Zhang, Junwei Zhang, Yanhong He. The Establishment and Optimization of a Regeneration System for Marigold (Tagetes erecta)[J]. Chinese Bulletin of Botany, 2020, 55(6): 749-759.
| No. | Genotype | Phenotype | Regeneration rate (%) | Source | Regeneration type | ||
|---|---|---|---|---|---|---|---|
| 1 | Perfect·Golden | Short-height plant, yellow-oran- ge color, double petal | 54.21±12.11 a | Chunhe Gardening (F1) | Easy | ||
| 2 | Milestone·Yellow | Short-height plant, yellow color, double petal | 51.44±8.29 ab | Sainuo Gardening (F1) | Easy | ||
| 3 | Proud·Orange-yellow | Short-height plant, yellow-oran- ge color, double petal | 41.47±6.12 abc | Dannuo Gardening (F1) | Moderate | ||
| 4 | Chrysanthemum marigold·Orange | Short-height plant, orange color, double petal | 36.23±11.95 abcd | Huaxianzi Gardening (F1) | Moderate | ||
| 5 | Xunlei·Yellow | Short-height plant, yellow color, double petal | 33.48±13.73 abcde | Chunhe Gardening (F1) | Moderate | ||
| 6 | Galore·Yellow | Medium-height plant, yellow co- lor, double petal | 31.95±18.22 abcdef | Hongyue Gardening (F1) | Moderate | ||
| 7 | Proud·Golden | Short-height plant, yellow-oran- ge color, double petal | 30.50±10.33 abcdef | Dannuo Gardening (F1) | Moderate | ||
| 8 | Double marigold· Light beige | Medium-height plant, yellow color, double petal | 22.01±14.56 bcdefg | Jinqiu Gardening (F1) | Difficult | ||
| 9 | DC | Short-height plant, orange color, double petal | 21.33±13.66 cdefgh | Self breeding (inbred line) * | Difficult | ||
| 10 | Youyi·Yellow | Short-height plant, yellow color, double petal | 18.00±1.83 cdefgh | Chunhe Gardening (F1) | Difficult | ||
| 11 | Miracle·Yellow | Short-height plant, yellow color, double petal | 13.29±9.62 cdefghi | Huaxianzi Gardening (F1) | Difficult | ||
| 12 | Old Inka·Orange | Medium-height plant, orange co- lor, double petal | 13.24±10.50 defghij | Hongyue Gardening (F1) | Difficult | ||
| 13 | Liberty·Yellow | Short-height plant, yellow color, double petal | 12.95±8.82 defghij | Huaxianzi Gardening (F1) | Difficult | ||
| 14 | 6210 | Medium-height plant, yellow-oran- ge color, double petal | 10.00±11.08 fghij | Self breeding (inbred line) * | Difficult | ||
| 15 | F8 | Medium-height plant, orange co- lor, simple petal | 9.33±5.96 efghij | Self breeding (inbred line) * | Difficult | ||
| No. | Genotype | Phenotype | Regeneration rate (%) | Source | Regeneration type | ||
| 16 | Fengsheng·Orange | Short-height plant, yellow-orange color, double petal | 7.11±7.70 ghij | Shijie seed (F1) | Difficult | ||
| 17 | AJ | Short-height plant, yellow-orange color, double petal | 7.02±13.41 ghij | Self breeding (inbred line) * | Difficult | ||
| 18 | Inka·Orange-yellow | Short-height plant, yellow-orange color, double petal | 5.11±7.87 ghij | Chunhe Gardening (F1) | Difficult | ||
| 19 | S3 | Medium-height plant, yellow color, simple petal | 4.92±2.75 ghij | Self breeding (inbred line) * | Difficult | ||
| 20 | S12 | Short-height plant, yellow color, simple petal | 4.67±10.43 hij | Self breeding (inbred line) * | Difficult | ||
| 21 | S5 | Short-height plant, yellow-orange color, simple petal | 4.67±5.06 ghij | Self breeding (inbred line) * | Difficult | ||
| 22 | SW2 | Medium-height plant, yellow color, double petal | 4.67±10.43 hij | Self breeding (inbred line) * | Difficult | ||
| 23 | Hawaii | Medium-height plant, yellow-oran- ge color, double petal | 4.22±5.87 ghij | Thailand (F1) | Difficult | ||
| 24 | 6170 | Medium-height plant, yellow-oran- ge color, double petal | 2.89±1.62 ghij | Self breeding (inbred line) * | Difficult | ||
| 25 | Jindun·Yellow | Short-height plant, yellow color, double petal | 2.71±4.36 hij | Sainuo Gardening (F1) | Difficult | ||
| 26 | GFR | Medium-height plant, orange color, double petal | 1.92±1.75 hij | Self breeding (inbred line) * | Difficult | ||
| 27 | DH | Short-height plant, yellow color, double petal | 1.33±1.83 ij | Self breeding (inbred line) * | Difficult | ||
| 28 | Vanilla | Medium-height plant, yellow-w- hite color, double petal | 0.91±2.03 ij | Thailand (F1) | Unable | ||
| 29 | Double Chrysanthe- mum marigold·Orange | Short-height plant, yellow-oran- ge color, double petal | 0.71±1.60 ij | Jinqiu Gardening (F1) | Unable | ||
| 30 | 6155 | Medium-height plant, yellow color, double petal | 0.00±0.00 j | Self breeding (inbred line) * | Unable | ||
| 31 | 9905 | Medium-height plant, yellow-oran- ge color, double petal | 0.00±0.00 j | Self breeding (inbred line) * | Unable | ||
| 32 | Antigua·Orange | Short-height plant, yellow-orange color, double petal | 0.00±0.00 j | Huaxianzi Gardening (F1) | Unable | ||
| 33 | Discovery·Orange | Short-height plant, orange color, double petal | 0.00±0.00 j | Sainuo Gardening (F1) | Unable | ||
| 34 | Double cut flower marigold·Lemon | Tall-height plant, yellow color, double petal | 0.00±0.00 j | Jinqiu Gardening (F1) | Unable | ||
| 35 | Double marigold·Golden | Medium-height plant, yellow color, double petal | 0.00±0.00 j | Jinqiu Gardening (F1) | Unable | ||
| 36 | Marigold·Deep orange | Tall-height plant, orange color, double petal | 0.00±0.00 j | Jinqiu Gardening (F1) | Unable | ||
| 37 | MB | Short-height plant, yellow color, double petal | 0.00±0.00 j | Self breeding (inbred line) * | Unable | ||
| 38 | New Fengsheng·Yellow | Short-height plant, yellow color, double petal | 0.00±0.00 j | Sainuo Gardening (F1) | Unable | ||
| 39 | S13 | Medium-height plant, yellow color, double petal | 0.00±0.00 j | Self breeding (inbred line) * | Unable | ||
| 40 | Yingtai·Golden | Medium-height plant, yellow color, double petal | 0.00±0.00 j | Shijie seed (F1) | Unable | ||
Table 1 Regeneration efficiency of different marigold genotypes (means±SD)
| No. | Genotype | Phenotype | Regeneration rate (%) | Source | Regeneration type | ||
|---|---|---|---|---|---|---|---|
| 1 | Perfect·Golden | Short-height plant, yellow-oran- ge color, double petal | 54.21±12.11 a | Chunhe Gardening (F1) | Easy | ||
| 2 | Milestone·Yellow | Short-height plant, yellow color, double petal | 51.44±8.29 ab | Sainuo Gardening (F1) | Easy | ||
| 3 | Proud·Orange-yellow | Short-height plant, yellow-oran- ge color, double petal | 41.47±6.12 abc | Dannuo Gardening (F1) | Moderate | ||
| 4 | Chrysanthemum marigold·Orange | Short-height plant, orange color, double petal | 36.23±11.95 abcd | Huaxianzi Gardening (F1) | Moderate | ||
| 5 | Xunlei·Yellow | Short-height plant, yellow color, double petal | 33.48±13.73 abcde | Chunhe Gardening (F1) | Moderate | ||
| 6 | Galore·Yellow | Medium-height plant, yellow co- lor, double petal | 31.95±18.22 abcdef | Hongyue Gardening (F1) | Moderate | ||
| 7 | Proud·Golden | Short-height plant, yellow-oran- ge color, double petal | 30.50±10.33 abcdef | Dannuo Gardening (F1) | Moderate | ||
| 8 | Double marigold· Light beige | Medium-height plant, yellow color, double petal | 22.01±14.56 bcdefg | Jinqiu Gardening (F1) | Difficult | ||
| 9 | DC | Short-height plant, orange color, double petal | 21.33±13.66 cdefgh | Self breeding (inbred line) * | Difficult | ||
| 10 | Youyi·Yellow | Short-height plant, yellow color, double petal | 18.00±1.83 cdefgh | Chunhe Gardening (F1) | Difficult | ||
| 11 | Miracle·Yellow | Short-height plant, yellow color, double petal | 13.29±9.62 cdefghi | Huaxianzi Gardening (F1) | Difficult | ||
| 12 | Old Inka·Orange | Medium-height plant, orange co- lor, double petal | 13.24±10.50 defghij | Hongyue Gardening (F1) | Difficult | ||
| 13 | Liberty·Yellow | Short-height plant, yellow color, double petal | 12.95±8.82 defghij | Huaxianzi Gardening (F1) | Difficult | ||
| 14 | 6210 | Medium-height plant, yellow-oran- ge color, double petal | 10.00±11.08 fghij | Self breeding (inbred line) * | Difficult | ||
| 15 | F8 | Medium-height plant, orange co- lor, simple petal | 9.33±5.96 efghij | Self breeding (inbred line) * | Difficult | ||
| No. | Genotype | Phenotype | Regeneration rate (%) | Source | Regeneration type | ||
| 16 | Fengsheng·Orange | Short-height plant, yellow-orange color, double petal | 7.11±7.70 ghij | Shijie seed (F1) | Difficult | ||
| 17 | AJ | Short-height plant, yellow-orange color, double petal | 7.02±13.41 ghij | Self breeding (inbred line) * | Difficult | ||
| 18 | Inka·Orange-yellow | Short-height plant, yellow-orange color, double petal | 5.11±7.87 ghij | Chunhe Gardening (F1) | Difficult | ||
| 19 | S3 | Medium-height plant, yellow color, simple petal | 4.92±2.75 ghij | Self breeding (inbred line) * | Difficult | ||
| 20 | S12 | Short-height plant, yellow color, simple petal | 4.67±10.43 hij | Self breeding (inbred line) * | Difficult | ||
| 21 | S5 | Short-height plant, yellow-orange color, simple petal | 4.67±5.06 ghij | Self breeding (inbred line) * | Difficult | ||
| 22 | SW2 | Medium-height plant, yellow color, double petal | 4.67±10.43 hij | Self breeding (inbred line) * | Difficult | ||
| 23 | Hawaii | Medium-height plant, yellow-oran- ge color, double petal | 4.22±5.87 ghij | Thailand (F1) | Difficult | ||
| 24 | 6170 | Medium-height plant, yellow-oran- ge color, double petal | 2.89±1.62 ghij | Self breeding (inbred line) * | Difficult | ||
| 25 | Jindun·Yellow | Short-height plant, yellow color, double petal | 2.71±4.36 hij | Sainuo Gardening (F1) | Difficult | ||
| 26 | GFR | Medium-height plant, orange color, double petal | 1.92±1.75 hij | Self breeding (inbred line) * | Difficult | ||
| 27 | DH | Short-height plant, yellow color, double petal | 1.33±1.83 ij | Self breeding (inbred line) * | Difficult | ||
| 28 | Vanilla | Medium-height plant, yellow-w- hite color, double petal | 0.91±2.03 ij | Thailand (F1) | Unable | ||
| 29 | Double Chrysanthe- mum marigold·Orange | Short-height plant, yellow-oran- ge color, double petal | 0.71±1.60 ij | Jinqiu Gardening (F1) | Unable | ||
| 30 | 6155 | Medium-height plant, yellow color, double petal | 0.00±0.00 j | Self breeding (inbred line) * | Unable | ||
| 31 | 9905 | Medium-height plant, yellow-oran- ge color, double petal | 0.00±0.00 j | Self breeding (inbred line) * | Unable | ||
| 32 | Antigua·Orange | Short-height plant, yellow-orange color, double petal | 0.00±0.00 j | Huaxianzi Gardening (F1) | Unable | ||
| 33 | Discovery·Orange | Short-height plant, orange color, double petal | 0.00±0.00 j | Sainuo Gardening (F1) | Unable | ||
| 34 | Double cut flower marigold·Lemon | Tall-height plant, yellow color, double petal | 0.00±0.00 j | Jinqiu Gardening (F1) | Unable | ||
| 35 | Double marigold·Golden | Medium-height plant, yellow color, double petal | 0.00±0.00 j | Jinqiu Gardening (F1) | Unable | ||
| 36 | Marigold·Deep orange | Tall-height plant, orange color, double petal | 0.00±0.00 j | Jinqiu Gardening (F1) | Unable | ||
| 37 | MB | Short-height plant, yellow color, double petal | 0.00±0.00 j | Self breeding (inbred line) * | Unable | ||
| 38 | New Fengsheng·Yellow | Short-height plant, yellow color, double petal | 0.00±0.00 j | Sainuo Gardening (F1) | Unable | ||
| 39 | S13 | Medium-height plant, yellow color, double petal | 0.00±0.00 j | Self breeding (inbred line) * | Unable | ||
| 40 | Yingtai·Golden | Medium-height plant, yellow color, double petal | 0.00±0.00 j | Shijie seed (F1) | Unable | ||
| No. | 6-BA (mg?L-1) | IAA (mg?L-1) | Regeneration rate (%) |
|---|---|---|---|
| 1 | 3 | 1.0 | 37.50±15.00 b |
| 2 | 3 | 1.5 | 40.00±14.14 b |
| 3 | 3 | 1.8 | 35.00±5.77 b |
| 4 | 3 | 2.0 | 52.50±5.00 ab |
| 5 | 3 | 2.5 | 47.50±17.08 ab |
| 6 | 3 | 3.0 | 65.00±12.91 a |
Table 2 Effect of 6-BA and IAA on leaf regeneration of marigold Milestone·Yellow
| No. | 6-BA (mg?L-1) | IAA (mg?L-1) | Regeneration rate (%) |
|---|---|---|---|
| 1 | 3 | 1.0 | 37.50±15.00 b |
| 2 | 3 | 1.5 | 40.00±14.14 b |
| 3 | 3 | 1.8 | 35.00±5.77 b |
| 4 | 3 | 2.0 | 52.50±5.00 ab |
| 5 | 3 | 2.5 | 47.50±17.08 ab |
| 6 | 3 | 3.0 | 65.00±12.91 a |
| No. | ZT (mg?L-1) | IAA (mg?L-1) | Regeneration rate (%) |
|---|---|---|---|
| 1 | 0.3 | 0.5 | 37.50±9.57 bc |
| 2 | 0.3 | 1.0 | 35.00±10.00 bc |
| 3 | 0.3 | 2.0 | 22.50±12.58 c |
| 4 | 0.6 | 0.5 | 67.50±9.57 a |
| 5 | 0.6 | 1.0 | 40.00±8.16 bc |
| 6 | 0.6 | 2.0 | 55.00±10.00 ab |
| 7 | 1.8 | 0.5 | 52.50±15.00 ab |
| 8 | 1.8 | 1.0 | 55.00±19.15 ab |
| 9 | 1.8 | 2.0 | 27.50±22.17 c |
Table 3 Effect of ZT and IAA on the leaf regeneration of marigold Milestone·Yellow
| No. | ZT (mg?L-1) | IAA (mg?L-1) | Regeneration rate (%) |
|---|---|---|---|
| 1 | 0.3 | 0.5 | 37.50±9.57 bc |
| 2 | 0.3 | 1.0 | 35.00±10.00 bc |
| 3 | 0.3 | 2.0 | 22.50±12.58 c |
| 4 | 0.6 | 0.5 | 67.50±9.57 a |
| 5 | 0.6 | 1.0 | 40.00±8.16 bc |
| 6 | 0.6 | 2.0 | 55.00±10.00 ab |
| 7 | 1.8 | 0.5 | 52.50±15.00 ab |
| 8 | 1.8 | 1.0 | 55.00±19.15 ab |
| 9 | 1.8 | 2.0 | 27.50±22.17 c |
| No. | TDZ (mg?L-1) | IBA (mg?L-1) | Regeneration rate (%) |
|---|---|---|---|
| 1 | 0.5 | 0.3 | 62.50±12.58 ab |
| 2 | 0.5 | 0.5 | 55.00±17.32 ab |
| 3 | 0.5 | 1.0 | 67.50±15.00 a |
| 4 | 1.0 | 0.3 | 40.00±14.14 bcd |
| 5 | 1.0 | 0.5 | 57.50±20.62 ab |
| 6 | 1.0 | 1.0 | 47.50±9.57 abcd |
| 7 | 1.5 | 0.3 | 32.50±12.58 cd |
| 8 | 1.5 | 0.5 | 52.50±9.57 abc |
| 9 | 1.5 | 1.0 | 27.50±9.57 d |
Table 4 Effect of TDZ and IBA on the leaf regeneration of marigold Milestone·Yellow
| No. | TDZ (mg?L-1) | IBA (mg?L-1) | Regeneration rate (%) |
|---|---|---|---|
| 1 | 0.5 | 0.3 | 62.50±12.58 ab |
| 2 | 0.5 | 0.5 | 55.00±17.32 ab |
| 3 | 0.5 | 1.0 | 67.50±15.00 a |
| 4 | 1.0 | 0.3 | 40.00±14.14 bcd |
| 5 | 1.0 | 0.5 | 57.50±20.62 ab |
| 6 | 1.0 | 1.0 | 47.50±9.57 abcd |
| 7 | 1.5 | 0.3 | 32.50±12.58 cd |
| 8 | 1.5 | 0.5 | 52.50±9.57 abc |
| 9 | 1.5 | 1.0 | 27.50±9.57 d |
| No. | TDZ (mg?L-1) | IBA (mg?L-1) | Regeneration rate (%) |
|---|---|---|---|
| 1 | 0.1 | 0.25 | 22.50±15.00 c |
| 2 | 0.1 | 0.50 | 40.00±8.16 ab |
| 3 | 0.2 | 0.25 | 35.00±12.91 ab |
| 4 | 0.2 | 0.50 | 62.50±12.58 a |
| 5 | 0.3 | 0.25 | 35.00±12.91 ab |
| 6 | 0.3 | 0.50 | 47.50±12.58 ab |
| 7 | 0.4 | 0.25 | 35.00±12.91 ab |
| 8 | 0.4 | 0.50 | 32.50±17.08 ab |
Table 5 Effect of TDZ and IBA on the leaf regeneration of marigold Milestone·Yellow
| No. | TDZ (mg?L-1) | IBA (mg?L-1) | Regeneration rate (%) |
|---|---|---|---|
| 1 | 0.1 | 0.25 | 22.50±15.00 c |
| 2 | 0.1 | 0.50 | 40.00±8.16 ab |
| 3 | 0.2 | 0.25 | 35.00±12.91 ab |
| 4 | 0.2 | 0.50 | 62.50±12.58 a |
| 5 | 0.3 | 0.25 | 35.00±12.91 ab |
| 6 | 0.3 | 0.50 | 47.50±12.58 ab |
| 7 | 0.4 | 0.25 | 35.00±12.91 ab |
| 8 | 0.4 | 0.50 | 32.50±17.08 ab |
| Code | Treatment | Explant No. | Elongation rate (%) | Status |
|---|---|---|---|---|
| MS1 | No hormone | 46 | 91.30 | Strong; elongate 4-5 cm |
| MS2 | 0.1 mg·L?1 TDZ; chang to 0.01 mg·L?1 TDZ after 15 days | 60 | 65.00 | Healthy; elongate 3-4 cm |
| MS3 | 0.3 mg·L?1 6-BA+1.0 mg·L?1 NAA | 53 | 30.19 | Weak; elongate 0-2 cm |
| MS4 | 0.6 mg·L?1 6-BA+1.0 mg·L?1 NAA | 55 | 18.18 | Weak; elongate 0-1 cm |
| MS5 | 0.9 mg·L?1 6-BA+1.0 mg·L?1 NAA | 57 | 12.28 | Weak; elongate 0-1 cm |
| MS6 | 0.2 mg·L?1 6-BA+0.6 mg·L?1 NAA | 45 | 33.33 | Weak; elongate 0-2 cm |
Table 7 Effects of different media on the elongation of marigold adventitious buds
| Code | Treatment | Explant No. | Elongation rate (%) | Status |
|---|---|---|---|---|
| MS1 | No hormone | 46 | 91.30 | Strong; elongate 4-5 cm |
| MS2 | 0.1 mg·L?1 TDZ; chang to 0.01 mg·L?1 TDZ after 15 days | 60 | 65.00 | Healthy; elongate 3-4 cm |
| MS3 | 0.3 mg·L?1 6-BA+1.0 mg·L?1 NAA | 53 | 30.19 | Weak; elongate 0-2 cm |
| MS4 | 0.6 mg·L?1 6-BA+1.0 mg·L?1 NAA | 55 | 18.18 | Weak; elongate 0-1 cm |
| MS5 | 0.9 mg·L?1 6-BA+1.0 mg·L?1 NAA | 57 | 12.28 | Weak; elongate 0-1 cm |
| MS6 | 0.2 mg·L?1 6-BA+0.6 mg·L?1 NAA | 45 | 33.33 | Weak; elongate 0-2 cm |
Figure 1 Leaves regenerations process of marigold (A) An easy regeneration genotype: Milestone·Yellow; (B) A moderate regeneration genotype: Proud·Orange-yellow; (C) A difficult regeneration genotype: Miracle·Yellow; (D) An unregenerable genotype: Antigua·Orange; (E) The leaves of Milestone·Yellow on the MS medium supplied with 3 mg·L?1 6-BA, 3 mg·L?1 IAA, 3 g·L?1 phytagel, and 30 g·L?1 sucrose; (F) The leaves of Milestone·Yellow on the MS medium supplied with 0.3 mg·L?1 ZT, 2.0 mg·L?1 IAA, 3 g·L?1 phytagel, and 30 g·L?1 sucrose; (G) The leaves of Milestone·Yellow on the MS medium supplied with 0.6 mg·L?1 ZT, 0.5 mg·L?1 IAA, 3 g·L?1 phytagel, and 30 g·L?1 sucrose; (H) The leaves of Milestone·Yellow on the MS medium supplied with 0.2 mg·L?1 TDZ, 0.5 mg·L?1 IBA, 3 g·L?1 phytagel, and 30 g·L?1 sucrose; (I) The leaves of Milestone·Yellow on the MS medium supplied with 0.2 mg·L?1 TDZ, 0.5 mg·L?1 IBA, 8 g·L?1 agar, and 30 g·L?1 sucrose; (J) The leaves of Milestone·Yellow on the MS medium supplied with 0.2 mg·L?1 TDZ, 0.5 mg·L?1 IBA, 8 g·L?1 agar, and 40 g·L?1 sucrose; (K) Regenerated shoot extension and rooting culture on MS1 medium; (L) A living regenerating transplanted plants. (A)-(K) Bars=1 cm; (L) Bar=2 cm
| Leaflet segment | Regeneration rate (%) | Status |
|---|---|---|
| Whole leaflet | 66.67±11.55 a | Callus and buds differentiated at the incision |
| Basal segment | 63.49±8.64 a | Callus were differentiated at both end incisions, but only adventitious buds were differentiated in basal wound |
| Top segment | 29.84±11.75 b | Callus and buds differentiated at the incision |
Table 6 Effect of lobular parts on the leaf regeneration of marigold
| Leaflet segment | Regeneration rate (%) | Status |
|---|---|---|
| Whole leaflet | 66.67±11.55 a | Callus and buds differentiated at the incision |
| Basal segment | 63.49±8.64 a | Callus were differentiated at both end incisions, but only adventitious buds were differentiated in basal wound |
| Top segment | 29.84±11.75 b | Callus and buds differentiated at the incision |
Figure 2 Effects of different curing agents and sucrose on the leaf regeneration and vitrification of marigold Milestone·Yellow (A), (B) Different types and concentrations of curing agent (T1: 3 g·L?1 phytagel; T2: 4.5 g·L?1 phytagel; T3: 6 g·L?1 phytagel; T4: 7 g·L?1 agar; T5: 8 g·L?1 agar; T6: 9 g·L?1 agar; T7: 10 g·L?1 agar); (C), (D) Different sucrose concentrations. Different lowercase letters indicate significant differences at P<0.05 level.
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