Silencing of omega-5 gliadins in transgenic wheat eliminates a major source of environmental variability and improves dough mixing properties of flour

Altenbach, Susan B; Tanaka, Charlene K; Seabourn, Bradford W

doi:10.1186/s12870-014-0393-1

Table 1 Changes in average normalized spot volumes of flour proteins in transgenic lines relative to the non-transgenic control grown with and without post-anthesis fertilizer

From: Silencing of omega-5 gliadins in transgenic wheat eliminates a major source of environmental variability and improves dough mixing properties of flour

		# 2-DE spots that decrease ¹				# 2-DE spots that increase ¹				Average spot volume ²						% Change ³
		Minus fertilizer		Plus fertilizer		Minus fertilizer		Plus fertilizer		Minus fertilizer			Plus fertilizer			Minus fertilizer		Plus fertilizer
# Spots ⁴	Protein Identification ⁵	35b	45a	35b	45a	35b	45a	35b	45a	Control	35b	45a	Control	35b	45a	35b	45a	35b	45a
8	HMW-GS Ax2^*									478.37	478.88	501.45	561.18	557.40	570.27	0.1	4.8	−0.7	1.6
8	HMW-GS Bx7									824.00	850.20	871.81	968.99	987.02	938.06	3.2	5.8	1.9	−3.2
8	HMW-GS By9						3		1	417.26	431.28	505.14	550.88	583.14	635.55	3.4	21.1	5.9	15.4
8	HMW-GS Dx5		3	2	4					755.94	722.52	652.04	848.64	822.21	684.83	−4.4	−13.7	−3.1	−19.3
7	HMW-GS Dy10						1		1	594.53	597.64	650.34	702.68	707.96	751.93	0.5	9.4	0.8	7.0
39	Total HMW-GS									3070.10	3080.52	3180.79	3632.37	3657.73	3580.64	0.3	3.6	0.7	−1.4
2	LMW-GS Bu-1 (m-type)⁶									625.10	600.98	646.79	647.11	629.87	653.32	−3.9	3.5	−2.7	1.0
4	LMW-GS Bu-2/13 (s-type)⁶				2					551.57	552.98	493.66	638.08	640.46	532.08	0.3	−10.5	0.4	−16.6
8	LMW-GS Bu-3 (s-type)⁶	1	4	1	3					1266.56	1275.58	1061.02	1264.48	1249.93	1048.88	0.7	−16.2	−1.2	−17.1
3	LMW-GS Bu-4 (i-type)⁶									353.30	380.21	403.80	377.56	380.24	405.58	7.6	4.3	0.7	7.4
1	LMW-GS Bu-6 (m-type)⁶									254.17	253.36	249.13	226.14	214.33	208.26	−0.3	−2.0	−5.2	−7.9
3	LMW-GS Bu-7 (m-type)⁶			1						523.29	534.42	572.33	499.02	497.83	521.43	2.1	9.4	−0.2	4.5
1	LMW-GS Bu-8 (m-type)⁶									32.70	31.85	28.07	37.40	31.53	37.11	−2.6	−14.2	−15.7	−0.8
1	LMW-GS Bu-11 (m-type)⁶									124.72	132.19	140.70	107.97	108.53	110.90	6.0	2.8	0.5	2.7
2	LMW-GS Bu-18 (m-type)⁶									270.35	293.55	326.37	255.90	265.262	56.40	8.6	*20.7*	3.7	0.2
1	LMW-GS TC11-277270 (m-type)⁶		1		1					150.06	150.51	149.04	149.07	167.17	138.69	0.3	−0.7	12.1	−7.0
1	LMW-GS [GenBank: AAB48469] (i-type)⁶									60.73	54.40	34.03	68.93	67.85	40.05	−10.4	−44.0	−1.6	−41.9
27	Total traditional LMW-GS									4212.57	4260.01	4104.93	4271.68	4253.01	3952.71	1.1	−2.6	−0.4	−7.5
2	Alpha chain-terminating gliadin Bu-2⁷						1		1	402.61	410.38	451.61	451.36	455.88	485.50	1.9	12.2	1.0	7.6
3	Gamma chain-terminating gliadin Bu-4⁸		2	1	1					476.24	481.89	415.65	473.78	478.97	430.01	1.2	−12.7	1.1	−9.2
4	Omega-1,2 chain-terminating gliadin⁹		3		4					283.38	277.98	195.36	536.24	537.38	246.83	−1.9	−31.1	0.2	−54.0
9	Total chain-terminating gliadins									1162.23	1170.26	1062.62	1461.38	1472.22	1162.33	0.7	−8.6	0.7	−20.5
1	Alpha gliadin Bu-1									68.09	68.56	74.33	81.37	69.73	85.50	0.7	9.2	−14.3	5.1
1	Alpha gliadin Bu-3									180.74	182.36	204.19	210.70	221.22	232.06	0.9	13.0	5.0	10.1
1	Alpha gliadin Bu-4									306.35	313.67	349.35	353.82	354.76	367.54	2.4	4.0	0.3	3.9
2	Alpha gliadin Bu-5						1			190.87	209.23	239.66	285.97	296.50	312.78	9.6	25.6	3.7	9.4
1	Alpha gliadin Bu-10									196.26	206.54	197.39	208.34	214.01	201.55	5.2	0.6	2.7	−3.3
1	Alpha gliadin Bu-11									205.24	212.00	245.84	272.06	279.76	303.77	3.3	19.8	2.8	11.7
4	Alpha gliadin Bu-12						1			506.20	511.08	617.13	698.95	707.77	784.63	1.0	21.9	1.3	12.3
4	Alpha gliadin Bu-14						2	1	3	271.13	289.98	348.01	358.25	384.61	446.39	7.0	28.4	7.4	24.6
1	Alpha gliadin Bu-23									290.26	297.86	277.91	336.09	340.86	330.65	2.6	−4.3	1.4	−1.6
1	Alpha gliadin Bu-27	1		1			1			114.68	50.80	155.69	142.19	41.25	169.12	−55.7	35.8	−71.0	18.9
1	Alpha gliadin Bu-BQ806209									80.78	81.85	73.91	74.03	66.24	3.0	4.4	0.2	−10.4	80.78
1	Alpha gliadin Bu-BQ807130									96.61	107.05	114.95	125.15	119.48	135.56	10.8	19.0	−4.5	8.3
19	Total alpha gliadin									2504.83	2529.92	2906.30	3146.79	3103.98	3435.79	1.0	16.0	−1.4	9.2
2	Gamma gliadin Bu-1		1		2					205.88	217.38	179.18	223.09	212.75	176.47	5.6	−13.0	−4.6	−20.9
1	Gamma gliadin Bu-2									298.32	306.62	266.99	337.45	332.72	295.21	2.8	−10.5	−1.4	−12.5
4	Gamma gliadin Bu-5		1							795.04	816.60	738.76	798.55	802.41	719.11	2.7	−7.1	0.5	−9.9
3	Gamma gliadin Bu-6				2					294.02	308.72	291.19	341.59	321.44	282.37	5.0	−1.0	−5.9	−17.3
1	Gamma gliadin Bu-7									109.93	115.62	119.32	117.19	110.12	126.22	5.2	8.5	−6.0	7.7
2	Gamma gliadin Bu-11									281.31	285.23	300.08	303.39	296.76	315.99	1.4	6.7	−2.2	4.2
13	Total gamma gliadin									1984.50	2050.18	1895.51	2121.27	2076.21	1915.37	3.3	−4.5	−2.1	−9.7
6	Omega-5 gliadin	6	6	6	6					675.82	229.83	146.27	1153.93	237.46	140.56	−66.0	−78.4	−79.4	−87.8
2	Omega-1,2 gliadin D3¹⁰		1		2					479.80	458.24	393.76	631.49	581.38	410.86	−4.5	−17.9	−7.9	−34.9
2	Omega-1,2 gliadin Bu-D5¹⁰	1	2	1	2					242.54	214.06	111.87	435.25	355.39	133.28	−11.7	−53.9	−18.3	−69.4
1	Secalin-like omega gliadin									37.73	41.88	49.08	49.63	52.13	56.00	11.0	*30.1*	5.1	12.8
11	Total omega gliadin									1435.89	944.01	700.98	2270.30	1226.37	740.69	−34.3	−51.2	−46.0	−67.4
4	Total mixed gliadin									404.96	425.58	424.86	432.88	460.34	472.80	51	4.9	6.3	9.2
7	Total triticin						1		3	673.65	727.12	794.69	777.11	839.56	964.18	7.9	18.0	8.0	24.1
1	Farinin Bu-1									58.89	55.48	68.66	41.31	33.45	39.08	−5.8	16.6	−19.0	−5.4
3	Farinin Bu-2									334.76	320.74	307.56	231.53	221.26	221.18	−4.2	−8.1	−4.4	−4.5
2	Farinin Bu-3									143.03	154.74	146.48	125.87	124.27	128.67	8.2	2.4	−1.3	2.2
6	Total farinin									536.68	530.95	522.70	398.71	378.98	388.94	−1.1	2.6	−4.9	2.5
2	Purinin Bu-1						1			204.59	217.28	239.48	180.15	162.16	175.04	6.2	17.1	−10.0	−2.8
2	Purinin Bu-2				1					213.30	224.44	247.53	185.90	174.15	168.17	5.2	16.0	−6.3	−9.5
2	Purinin Bu-3									457.02	475.77	529.57	423.85	423.25	400.29	4.1	15.9	−0.1	−5.6
6	Total purinin									874.92	917.50	1016.58	789.89	759.56	743.49	4.9	16.2	−3.8	−5.9
1	Globulin-1									41.35	41.57	52.98	58.73	58.46	70.51	0.5	*28.1*	−0.5	*20.1*
5	Globulin-2	1	2	2	2					115.61	105.93	102.16	104.84	98.49	91.10	−8.4	−11.6	−6.1	−13.1
4	Globulin Glo-3		1		1					246.03	257.17	215.45	210.61	225.19	187.46	4.5	−12.4	6.9	−11.0
10	Total globulin									402.98	404.67	370.59	374.18	382.14	349.08	0.4	−8.0	2.1	−6.7
4	Serpin Bu-1						3			168.10	162.31	211.52	251.48	253.04	278.30	−3.4	25.8	0.6	10.7
2	Serpin Bu-2						2			53.48	52.28	70.47	108.94	114.09	119.34	−2.2	31.8	4.7	9.6
2	Serpin Bu-3								1	41.23	40.67	50.63	63.64	66.61	81.15	−1.4	22.8	4.7	27.5
1	Serpin Bu-4						1			67.65	69.35	83.86	105.73	111.12	128.48	2.5	*24.0*	5.1	*21.5*
2	Serpin Bu-5									83.63	83.32	103.75	126.05	125.89	152.76	−0.4	24.1	−0.1	21.2
2	Serpin Bu-7	1	1	1	1	1	1	1	1	59.27	62.64	63.17	74.88	70.10	73.42	5.7	6.6	−6.4	−2.0
13	Total serpin									473.36	470.56	583.41	730.71	740.86	833.45	−0.6	23.2	1.4	14.1
2	Beta-amylase Bu-2									102.36	93.67	101.60	99.71	98.17	99.37	−8.5	−0.7	−1.5	−0.3
3	Beta-amylase Bu-3	1	2	3	1					117.39	99.37	100.84	136.55	87.79	98.43	−15.4	−14.1	−35.7	−27.9
5	Total beta-amylase									219.75	193.03	202.43	236.27	185.97	197.80	−12.2	−7.9	−21.3	−16.3
2	WMAI Bu-1									624.43	660.27	661.82	469.28	488.91	456.32	5.7	6.0	4.2	−2.8
1	WDAI Bu-1									616.66	621.14	659.08	506.27	464.64	489.27	0.7	6.9	−8.2	−3.4
1	WDAI Bu-3									256.36	259.57	294.40	245.10	230.20	273.67	1.3	14.8	−6.1	11.7
1	WDAI Bu-4									179.79	194.48	206.86	130.61	113.52	126.49	8.2	15.1	−13.1	−3.1
1	WTAI CM1									223.52	233.25	247.31	183.44	172.34	170.34	4.3	10.6	−6.1	−7.1
2	WTAI CM2			1						1028.76	1046.46	1023.65	760.84	682.30	707.44	1.7	−0.5	−10.3	−7.0
2	WTAI CM3									653.13	660.93	658.76	483.08	436.22	402.44	1.2	0.9	−9.7	−16.7
2	WTAI CM16			1	1					549.43	517.83	535.74	397.30	332.88	337.40	−5.8	−2.5	−16.2	−15.1
2	WTAI CM17									442.72	437.00	443.39	307.31	263.48	266.70	−1.3	0.1	−14.3	−13.2
2	WCI									402.00	420.67	403.68	281.27	260.35	243.91	4.6	0.4	−7.4	−13.3
2	CMX1/CMX3									256.24	268.07	268.12	197.36	191.19	184.98	4.6	4.6	−3.1	−6.3
1	WASI									250.63	248.99	262.19	177.70	184.67	188.80	−0.7	4.6	3.9	6.2
19	Total amylase/protease inhibitors									5483.67	5568.66	5664.99	4139.56	3820.69	3847.76	1.5	3.3	−7.7	−7.0
32	Other identified proteins		1		1	1				2056.00	2108.86	2064.67	1888.87	1871.32	1966.62	2.6	0.4	−0.9	4.1

¹Number of spots identified as each protein type that increased or decreased at least 20% in transgenic lines relative to non-transgenic control. Spot numbers and identities of all 2-DE spots that change can be found in Additional file 3.
²Sum of average volumes of all spots with the same MS/MS identification. MS/MS identifications are from [6].
³Protein types that that increased or decreased at least 20% in transgenic lines relative to non-transgenic control are indicated in bold. Changes that were greater than 20% but were not deemed to be significant by ANOVA are shown in bold italics.
⁴Total number of spots in 2-D gels that were identified as the same protein type in [6].
⁵Predominant protein in spots identified by MS/MS in [6]. Protein classes are indicated in bold type.
⁶Classification of traditional LMW-GS as m-type, s-type and i-type is based on the first amino acid of the mature proteins; methionine, serine or isoleucine.
⁷Protein has amino acid sequence similar to an alpha gliadin but contains seven cysteines that enables it to be linked into the glutenin polymer.
⁸Protein has amino acid sequence similar to a gamma gliadin but contains nine cysteines that enables it to be linked into the glutenin polymer.
⁹Protein has amino acid sequence similar to an omega-1,2 gliadin but contains one cysteine that enables it to be linked into the glutenin polymer.
¹⁰Two distinct protein types were distinguished for omega-1,2 gliadins. Percent changes of total omega-1,2 gliadins are −6.9 and −30.0 for 35b and 45a grown without fertilizer, respectively, and −12.2 and −49.0 for 35b and 45a grown with fertilizer, respectively.

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