Bulletin of Botanical Research ›› 2023, Vol. 43 ›› Issue (3): 351-360.doi: 10.7525/j.issn.1673-5102.2023.03.004
• Genetic and Breeding • Previous Articles Next Articles
Li CAO1, Yunli YANG1, Tianfang LI2, Jing JIANG1(
)
Received:2022-10-04
Online:2023-05-20
Published:2023-05-11
Contact:
Jing JIANG
E-mail:jiangjing1960@126.com
About author:CAO Li（1998—），female，postgraduate，research on genetic breeding of forest trees.
Supported by:CLC Number:
Li CAO, Yunli YANG, Tianfang LI, Jing JIANG. Analysis on Leaf Color and Growth Variation of Transgenic BpGLK Betula pendula ‘Dalecarlica'[J]. Bulletin of Botanical Research, 2023, 43(3): 351-360.
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URL: https://bbr.nefu.edu.cn/EN/10.7525/j.issn.1673-5102.2023.03.004
Table 1
Primer sequences
引物 Primer | 序列（5′→3′） Sequence（5′→3′） | 扩增片段长度 Amplified fragment length/bp |
|---|---|---|
| pFGC5941_CIS_F | CATGCCATGGGCACAGAAGGTTTGTGCAAG | 414 |
| pFGC5941_CIS_R | TTGGCGCGCCCCATACATCTGCCTTCTCTGG | |
| pFGC5941_Anti_F | GCTCTAGAGCACAGAAGGTTTGTGCAAG | 429 |
| pFGC5941_Anti_R | CGCGGATCCCCATACATCTGCCTTCTCTGG | |
| Bar-F | TTAGATCTCGGTGACGGGCA | 500 |
| Bar-R | CGGTCTGCACCATCGTCAAC |
Table 2
qRT-PCR primer sequence
引物 Primer | 引物序列（5′→3′） Sequence（5′→3′） |
|---|---|
| Bp18S-F | ATCTTGGGTTGGGCAGATCG |
| Bp18S-R | CATTACTC CGATCCCGAAGG |
| q-glk-F | CACAACATAGCCAGCCACCTTC |
| q-glk-R | GTCGGTGCTACCCAAGGACTC |
Fig.1
Electrophoresis pattern ofPCR amplification of BpGLK suppressed expression transgenic linesA.PCR for forward fragment；B.PCR for reverse fragment；C.PCR for Bar；M.DNA Maker DL2000；1.Positive plasmid；2.Negative control（ddH2O）；3.WT line；4-11.Transgenic lines RE1-RE8
Fig.2
Relative expression levels of BpGLK in transgenic plantletsDifferent lowercase letters indicate significant difference in relative expression among different lines at the same time（P<0.05）
Table 3
Temporal variation of L*value and b*value in tested lines
指标 Indexes | 株系 Lines | 5月15日 May 15th | 6月15日 Jun. 15th | 7月15日 Jul. 15th | 9月15日 Sep. 15th |
|---|---|---|---|---|---|
L*值 L*-value | WT | 37.42±0.42c | 40.41±0.50d | 37.36±0.49c | 36.68±0.66b |
| RE1 | 43.35±0.64a | 47.10±0.36a | 47.15±0.47a | 42.42±1.01a | |
| RE2 | 43.02±0.35a | 44.19±0.37b | 44.07±1.06ab | 42.30±1.13a | |
| RE3 | 44.11±0.63a | 44.43±0.60b | 40.78±2.66b | 43.35±1.20a | |
| RE4 | 42.23±0.69a | 43.21±0.54bc | 43.59±0.79b | 41.02±0.96a | |
| RE5 | 42.00±0.54a | 42.06±0.33c | 41.64±0.99b | 42.71±1.05a | |
| RE6 | 38.71±0.28bc | 36.85±0.76f | 36.90±0.71c | 38.03±0.63b | |
| RE7 | 42.87±1.51a | 39.26±0.41de | 36.43±0.98c | 37.60±0.83b | |
| RE8 | 41.35±2.57ab | 38.19±0.59ef | 36.54±0.80c | 35.87±1.03b | |
b*值 b*-value | WT | 19.27±0.64d | 19.86±0.81e | 19.63±0.64d | 16.26±1.08cd |
| RE1 | 29.38±1.23b | 31.72±0.61a | 35.40±0.69a | 26.16±1.37ab | |
| RE2 | 29.07±0.78bc | 27.96±0.38bc | 33.50±1.04a | 27.99±1.39a | |
| RE3 | 31.72±0.48a | 29.72±0.78ab | 35.41±0.40a | 28.48±0.94a | |
| RE4 | 27.18±0.78c | 26.97±0.86cd | 30.29±1.01b | 23.96±1.93b | |
| RE5 | 27.10±0.80c | 25.73±0.42d | 27.12±1.67c | 26.18±1.54ab | |
| RE6 | 20.64±0.47d | 18.90±1.08ef | 20.46±0.99d | 18.17±1.16cd | |
| RE7 | 20.91±0.37d | 20.64±0.52e | 20.19±0.99d | 18.82±1.26c | |
| RE8 | 21.43±0.58d | 17.59±0.67f | 20.36±0.71d | 14.79±0.69d |
Fig.3
Leaf color comparison of the tested lines
Table 4
Temporal variation of relative chlorophyll content（SPAD） in the tested lines
株系 Lines | 5月15日 May 15th | 6月15日 Jun. 15th | 7月15日 Jul. 15th | 9月15日 Sep. 15th |
|---|---|---|---|---|
| WT | 42.60±0.97a | 31.94±0.63a | 34.94±0.63bc | 39.27±1.28a |
| RE1 | 37.09±1.75bc | 23.97±1.02d | 31.37±0.90de | 27.03±1.36c |
| RE2 | 33.55±0.93cd | 26.60±1.47cd | 31.73±1.09de | 26.78±0.73c |
| RE3 | 32.27±0.78d | 24.41±0.71d | 30.65±0.81e | 27.88±1.08c |
| RE4 | 35.41±0.92cd | 24.77±0.65d | 34.14±0.96cd | 28.30±1.15c |
| RE5 | 40.68±0.93ab | 25.60±0.67d | 33.57±0.60cde | 22.36±0.76d |
| RE6 | 41.49±1.54a | 29.76±0.71ab | 39.19±1.70a | 39.87±2.39a |
| RE7 | 42.87±1.51a | 28.72±0.95bc | 37.51±0.83ab | 36.43±1.02ab |
| RE8 | 41.35±2.57a | 28.32±0.75bc | 36.05±0.77bc | 35.38±1.29b |
Table 5
Comparison of Chlorophyll a and Chlorophyll b mass fractions in the leaves of the tested lines
株系 Lines | 叶绿素a Chlorophyll a/（mg·g-1） | 叶绿素b Chlorophyll b/（mg·g-1） | 叶绿素a与叶绿素b比值 Ratio of Chlorophyll a to Chlorophyll b | ||||||
|---|---|---|---|---|---|---|---|---|---|
6月15日 Jun. 15th | 7月15日 Jul. 15th | 9月15日 Sep. 15th | 6月15日 Jun. 15th | 7月15日 Jul. 15th | 9月15日 Sep. 15th | 6月15日 Jun. 15th | 7月15日 Jul. 15th | 9月15日 Sep. 15th | |
| WT | 2.81±0.00ab | 2.72±0.03a | 2.81±0.00b | 1.36±0.01b | 1.22±0.04a | 1.29±0.00c | 2.07±0.02c | 2.23±0.05d | 2.19±0.00c |
| RE1 | 1.36±0.01f | 1.58±0.01g | 2.08±0.03e | 0.19±0.00e | 0.44±0.01f | 0.67±0.03e | 7.27±0.08a | 3.56±0.01b | 3.10±0.07a |
| RE2 | 1.66±0.01e | 1.75±0.03e | 2.66±0.02c | 0.24±0.04e | 0.68±0.06d | 1.00±0.02d | 7.24±1.26a | 2.63±0.20c | 2.67±0.04b |
| RE3 | 1.99±0.01d | 1.36±0.02h | 2.06±0.00e | 0.76±0.03c | 0.31±0.01g | 0.63±0.01e | 2.63±0.10bc | 4.43±0.15a | 3.25±0.03a |
| RE4 | 2.08±0.05cd | 1.93±0.01d | 2.55±0.02d | 0.73±0.13c | 0.54±0.00e | 0.96±0.01d | 3.08±0.61bc | 3.58±0.00b | 2.65±0.02b |
| RE5 | 2.17±0.10c | 1.64±0.01f | 2.64±0.05c | 0.54±0.04d | 0.38±0.01fg | 1.20±0.14c | 4.03±0.11b | 4.31±0.09a | 2.25±0.20c |
| RE6 | 2.91±0.01a | 2.17±0.01c | 2.65±0.05c | 1.68±0.03a | 0.86±0.01c | 1.58±0.11b | 1.74±0.04c | 2.51±0.03cd | 1.70±0.15d |
| RE7 | 2.71±0.06b | 2.63±0.04b | 2.98±0.01a | 1.26±0.08b | 1.08±0.04b | 1.51±0.01b | 2.16±0.09c | 2.44±0.12cd | 1.98±0.01c |
| RE8 | 2.79±0.03ab | 2.71±0.01a | 3.00±0.00a | 1.32±0.07b | 1.17±0.02a | 2.03±0.03a | 2.13±0.15c | 2.32±0.02cd | 1.48±0.02d |
Table 6
Comparison of total Chlorophyll and Carotenoids mass fractions in the leaves of the tested lines
株系 Lines | 总叶绿素 Total Chlorophyll/（mg·g-1） | 类胡萝卜素 Carotenoids/（mg·g-1） | ||||
|---|---|---|---|---|---|---|
6月15日 Jun. 15th | 7月15日 Jul. 15th | 9月15日 Sep. 15th | 6月15日 Jun. 15th | 7月15日 Jul. 15th | 9月15日 Sep. 15th | |
| WT | 4.17±0.01b | 3.94±0.07a | 4.10±0.00c | 0.41±0.00abc | 0.43±0.01b | 0.48±0.00ab |
| RE1 | 1.55±0.01e | 2.02±0.02e | 2.75±0.06f | 0.35±0.00cd | 0.32±0.00e | 0.41±0.00bc |
| RE2 | 1.90±0.04d | 2.42±0.09d | 3.66±0.04de | 0.42±0.02ab | 0.32±0.01e | 0.53±0.00a |
| RE3 | 2.75±0.03c | 1.67±0.03f | 2.70±0.01f | 0.35±0.01cd | 0.32±0.00e | 0.45±0.00bc |
| RE4 | 2.81±0.18c | 2.47±0.01d | 3.51±0.03e | 0.36±0.04bcd | 0.35±0.01d | 0.45±0.00bc |
| RE5 | 2.71±0.14c | 2.02±0.01e | 3.83±0.19d | 0.46±0.01a | 0.37±0.00c | 0.39±0.04c |
| RE6 | 4.60±0.02a | 3.03±0.02c | 4.23±0.10c | 0.33±0.01d | 0.39±0.00c | 0.32±0.06d |
| RE7 | 3.97±0.13b | 3.72±0.01b | 4.48±0.02b | 0.42±0.02ab | 0.43±0.01b | 0.46±0.00abc |
| RE8 | 4.11±0.05b | 3.88±0.02a | 5.04±0.03a | 0.39±0.03bcd | 0.46±0.01a | 0.28±0.01d |
Fig.4
Logistic fit curve of tree height of transgenic lines
Table 7
Logistic model of seedling height in transgenic birch
株系 Lines | 封顶后苗高实测值 Measured seedling height after capping/cm | R2 | 苗木开始生长时苗高 Seedling height when it starts to grow/cm | 苗木停止生长时苗高 Seedling height when it ceases to grow/cm | 速生点（t0） Fast growth point/d | 苗高速生点处生长速度 Seedling growth at high growth point/（cm·d-1） |
|---|---|---|---|---|---|---|
| WT | 247.00c | 0.998 | 105.27 | 247.00 | 171 | 1.698 |
| RE1 | 244.79c | 0.998 | 123.60 | 244.79 | 175 | 1.442 |
| RE2 | 245.93bc | 0.996 | 122.73 | 245.93 | 175 | 1.393 |
| RE3 | 215.71d | 0.999 | 112.87 | 215.71 | 177 | 1.353 |
| RE4 | 281.03ab | 0.984 | 113.51 | 281.03 | 182 | 1.602 |
| RE5 | 269.92c | 0.998 | 109.53 | 269.92 | 178 | 2.102 |
| RE6 | 285.71a | 0.999 | 119.33 | 285.71 | 179 | 2.111 |
| RE7 | 271.55bc | 0.997 | 115.00 | 271.55 | 175 | 1.805 |
| RE8 | 259.45c | 0.998 | 99.00 | 259.45 | 177 | 2.260 |
Table 8
Comparison of growth parameters of transgenic lines in the fast growth period
株系 Lines | 速生点（t0） Fast growth point/d | 速生持续时长 Duration time/d | 生长期参数Parameters in the growth stage | ||
|---|---|---|---|---|---|
苗高平均生长量（GR） Average growth of seedling height/cm | 苗高日生长量均值（GD） Mean daily growth of seedlings height/（cm·d-1） | 速生期生长量比率 Biomass ratio of fast growth period/% | |||
| WT | 171 | 59 | 87.911 | 1.490 | 56.56 |
| RE1 | 175 | 65 | 81.786 | 1.258 | 57.42 |
| RE2 | 175 | 74 | 90.193 | 1.219 | 56.80 |
| RE3 | 177 | 53 | 62.827 | 1.185 | 57.02 |
| RE4 | 182 | 75 | 104.902 | 1.399 | 57.23 |
| RE5 | 178 | 53 | 97.511 | 1.840 | 57.41 |
| RE6 | 179 | 54 | 100.060 | 1.853 | 56.81 |
| RE7 | 175 | 60 | 94.761 | 1.579 | 57.23 |
| RE8 | 177 | 49 | 96.984 | 1.979 | 57.36 |
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