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Metabolic manipulation of plants to improve their nutritional quality is an important …


Biology Articles » Biotechnology » Green Biotechnology » High-level tryptophan accumulation in seeds of transgenic rice and its limited effects on agronomic traits and seed metabolite profile » Tables

Tables
- High-level tryptophan accumulation in seeds of transgenic rice and its limited effects on agronomic traits and seed metabolite profile

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Table 1 Spikelet fertility of OASA1D transgenic plants (R0) grown in a greenhouse

Transgene Trp content Spikelet fertility (%)
Total no. of plants
≤20 21–40 41–60 61–80 81–100 Mean ±SD
OASA1D Increased 25 19 12 7 0 30.7±21.2 63
OASA2 Normal 5 4 6 14 11 61.4±27.2 40
NBa Normal 0 1 2 3 6 76.2±19.7 12

a Seed grown Nipponbare plants.

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Table 2 Total and free Trp contents of seeds from OASA1D transgenic plants grown in a greenhouse

Transgenic line Seed generation Seed weight (mg) Total Trp (nmol g–1) Free Trp (nmol g–1) Means ±SD
Nipponbare 18.6 4188 55.0±21.7
GUS16a R2 14.2 5173 41.0±8.0
M25 R2 11.7 8100
M121 R3 14.9 8810 3036.6±233.4
M31 R2 17.1 9237
M21 R2 9.9 9264
H29 R2 14.1 11 320
H36 R2 17.0 14 425 7642.3±830.3
H17 R3 16.2 14 178 12 309.0±5764.9
H37 R2 14.0 15 619
H42 R2 11.9 16 967
H40 R2 12.7 17 322
M45 R2 12.7 20 830 14 495.3±3528.0
M34 R2 7.7 48 519 23 705.3±11 681.2

Five seeds of one panicle from each transgenic line were used for measurement of total Trp content and of average seed weight. Total Trp content and seed weight of Nipponbare were determined as average values for two plants. Free Trp was measured in three to five single seeds, and values presented are means ±SD. Trp content is shown as nmol Trp g–1 dry seed weight.

a Transgenic line harbouring GUS and hph (OASA1D lines contain OASA1D and hph).

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Table 3 Nitrogen and Trp contents of seeds produced by HW lines under field or greenhouse conditions

Growth condition Line N content (%) Free Trp
Total Trp
(nmol g–1) R.V.a (nmol g–1) R.V.a
Greenhouse HW 1 1.39±0.12 (2) 2403±283.4 a (6) 55.9 7883±151.2 a (3) 1.9
HW 5 1.30±0.10 (2) 1762±333.3 b (6) 41.0 6949±387.6 b (3) 1.7
NB 1.24±0.04 (2) 43±15.7 c (6) 1 4118±237.0 c (3) 1
Field HW1 1.13±0.03 (2) 5294±2672.6 a (6) 311.0 8477±1139.0 a (4) 14.1
HW5 1.04±0.01 (2) 3288±1366.1 a (6) 193.0 5684±1818.0 b (4) 9.4
NB 0.96±0.13 (2) 17±9.4 b (9) 1 602±413.0 c (3) 1

Five seeds of Nipponbare (NB) or HW lines were analysed for nitrogen content (percentage of dry seed weight) and total Trp content (nmol g–1 dry seed weight) in each of the indicated (in parentheses) number of experiments, and data presented are means ±SD. Values for free Trp content (nmol g–1 dry seed weight) are presented as means ±SD for the numbers of seeds indicated in parentheses. Means followed by the same letters are not significantly different at the 5% level (Student's t test).

a Relative value compared with Nipponbare.

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Table 4 Free amino acid composition of seeds produced by field-grown HW lines

Amino acid Free amino acid content (nmol g–1)
NB
HW1
HW5
Mean ±SD (%)a Mean ±SD (%)a Relative valueb Mean ±SD (%)a Relative valueb
Gly 51±29 a 1.9 153±95 b 1.4 3.0 142±26 b 1.8 2.8
Ala 154±95 a 5.8 794±581 b 7.3 5.2 608±315 b 7.9 3.9
Val 32±19 a 1.2 84±37 b 0.8 2.6 68±16 b 0.9 2.1
Leu 13±7 a 0.5 36±20 b 0.3 2.7 32±8 b 0.4 2.5
Ile 8±5 a 0.3 28±15 b 0.3 3.5 26±9 b 0.3 3.3
Ser 172±95 a 6.5 288±107 a 2.6 1.7 277±68 a 3.6 1.6
Thr 37±30 a 1.4 70±37 a 0.6 1.9 57±31 a 0.7 1.5
Met 5±4 a 0.2 8±2 a 0.1 1.6 6±2 a 0.1 1.2
Asp 575±333 a 21.9 1285±727 a, b 11.7 2.2 1136±139 b 14.7 2.0
Asn 595±760 a 22.6 1269±616 a 11.6 2.1 746±296 a 9.7 1.3
Glu 660±359 a 25.1 810±303 a 7.4 1.2 739±165 a 9.6 1.1
Gln 51±31 a 1.9 47±13 a 0.4 0.9 54±8 a 0.7 1.1
Arg 67±71 a 2.5 239±123 b 2.2 3.6 102±48 a 1.3 1.5
Lys 16±7 a 0.6 65±34 b 0.6 4.1 48±14 b 0.6 3.0
His 28±17 a 1.1 115±48 b 1.1 4.1 116±75 b 1.5 4.1
Phe 7±4 a 0.3 30±15 b 0.3 4.3 25±7 b 0.3 3.6
Tyr 91±55 a 3.5 208±74 b 1.9 2.3 171±28 b 2.2 1.9
Trp 20±9 a 0.8 5294±2673 b 48.4 264.7 3288±1366 b 42.5 164.4
Pro 49±31 a 1.9 125±60 b 1.2 2.6 96±17 b 1.2 2.0
Total 2629±2001 a 100 10 947±4484 b 100 4.2 7734±1810 b 100 2.9

Data are expressed as nmol of amino acid g–1 dry seed weight and are means ±SD of values from six seeds of Nipponbare (NB) or HW lines. Means followed by the same letters are not significantly different at the 5% level (Student's t test).

a Amount of each amino acid as a percentage of total amino acids.

b Amount of each amino acid relative to that in Nipponbare.

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Table 5 Agronomic traits of HW1 and HW5 lines

Growth condition (year) Line na Days to heading ±SD Culm length (cm) Panicle length (cm) No. of tillers plant–1 Spikelet fertility (%) ±SDb No. of spikelets panicle–1 Mean dry matter of harvested seeds (30 plants g–1) ±SDc Pollen fertility (%)d Brown seed weight (1000 grains g–1)e
Greenhouse (2002) HW1 10 129±0.64 78.1 a 16.1 a 13.7 a 79.5±6.84 98.3 20.1 a
HW5 10 132±2.04 74.9 a 15.7 a 13.1 a 88.3±5.19 97.5 20.1 a
NB 23 130±1.13 80.1 a 17.0 a 11.6 a 88.2±6.65 98.9 20.1 a
Field (2003) HW1 2 100±0.00 73.3 b 18.1 a 15.5 a 65.0±18.21 71.6 b 443.6±35.64 97.6 20.2 ab
HW5 2 101±0.50 75.7 ab 17.8 a 13.2 a 81.3±11.00 85.6 b 588.4±50.77 98.5 21.0 a
NB 2 100±0.00 81.3 a 20.1 a 12.2 a 91.6±2.91 122.3 a 844.8±27.86 99.4 20.0 b

Means followed by the same letters are not significantly different at the 5% level (Student's t test).

an means repetition. The number of plants or plots for the greenhouse and field conditions, respectively. Heading date, culm length, panicle length, and the number of tillers were recorded for each of 10 or 23 plants in the greenhouse condition and for each of 15 plants per plot in the field condition.

b Spikelet fertility was recorded for all panicles of 10 plants in 2002 and for three panicles of main culms of 60 plants in 2003.

c Mean dry matter of harvested seeds which were air-dried (assumed moisture content of seeds was approximately 10–15%).

d Pollen fertility was recorded for three spikelets of each of three randomly selected plants. It was estimated from microscopic analysis of the shape and color of the pollen stained with I-KI solution (1%).

e The weight of 1000 seeds was determined from that of 20 or 100 grains in the greenhouse and field conditions, respectively.

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Table 6 Germination percentage and time to seedling establishment for HW lines

Line Greenhouse
Field
Germination (%) Time to establish seedlings (d) Germination (%) Time to establish seedlings (d)
HW1 95 6.3 53.3 10.1
HW5 100 3.1 93.3 5.3
NB 100 3 100 2.8

The germination test was performed with 20 seeds from greenhouse-grown plants and 15 seeds from field-grown plants. Data are means of values from two experiments.

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