Carbon assimilation and partitioning in darkness

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Read.Me Final Data Labelling Harvest Mass 18 Days 28 days

Carbon assimilation and partitioning, measured using 14CO2
Genotypes Col, prr7prr9; Ws, lhycca1
Labelling in darkness to test C fixation by PEPC without photosynthesis
Labelling protocol	From 'Methods' - PEPC activity and incorporation of labelled 14CO2 into metabolite pools was measured as previously described for whole-plant labelling (Kölling et al. 2013, Kölling et al. 2015) with minor modifications. Near the end of the night (ZT20-21), 28-day-old plants were transferred to a custom built, sealed chamber in the dark. Each replicate consisted of the aerial parts of a single plant. Labelling was initiated through the release of 14CO2 (150 μCi) from NaH14CO3 with the addition of lactic acid. After 1 h, the plants were harvested into 5 ml 80% (v/v) ethanol and incubated for 15 min at 80°C. Following homogenisation, the soluble fraction was collected by centrifugation (2,400g, 12 min), pooled with four sequential 1-ml washes (50% ethanol, 20% ethanol, water, and then 80% ethanol) of the insoluble fraction pellet. The soluble fraction was concentrated under vacuum. A water-soluble sub-fraction was collected by dissolving the near-dry exsiccate in 2 ml water, while the remainder was dissolved in 2 ml 98% ethanol. Basic, acidic, and neutral fractions were separated from the water-soluble fraction by ion-exchange chromatography as described (Quick et al. 1989). Partitioning into protein was measured as described (Kölling et al. 2013). Isotope incorporation into each fraction was measured by liquid scintillation counting.

Sheets in this workbook	Contents
Final Data	mean data, SD and charts
Labelling	Summary of experiment, replicates and timing (ZT)
Harvest Mass	Data on mass of harvested tissue, per replicate
18 days	Partitioning of 14C label in plants at 18 days - use these data for comparison of lhycca1 to its parent Ws
28 days	Partitioning of 14C label in plants at 28 days - note the lhycca1 genotype had started flowering at this timepoint

Credits
Experimental design and analysis	Gavin George, Sam Zeeman group at ETH Zurich
Curation	Andrew Millar



	18 Days
		EtOH				Water				Neutral			Acid				Basic				Insol				Starch				Protein				Cell wall				Assimilation
	Col-0	0.44783639721781787	±	0.04543542414093131		86.59258816279475	±	0.8032073167483322		0.08771570416643622	±	0.07048602777232704	40.52926432957474	±	2.5262081546687742		43.74125700701987	±	3.5226742845625796		12.959575439987427	±	0.8369099758029841		0.9177178972887473	±	0.7978715645510204		8.785055784169613	±	0.7612789559370693		3.256801758529068	±	0.4211181412095605		377.7537984343283	±	26.55853220216979
	prr7/9	0.7886351792247885	±	0.4024416923388231		82.5043008714346	±	0.8754554070053538	*	0.19421013151321795	±	0.13739603188905347	44.00025207583705	±	0.5584576389951075		34.65610012126357	±	1.4587549861856766	*	16.707063949340608	±	1.2604142979692203	*	0.03431775909206191	±	0.1617962073106456		13.719813498010012	±	1.0717337605792359	*	2.9529326922385333	±	0.42253706955437775		275.6563134032681	±	41.8160822643151	*
	WS	0.44192273868188675	±	0.03291544537103846		85.87199341501356	±	1.9861423039490287		-0.046192116560937246	±	0.10293146401224013	42.88782457399689	±	2.168316353512296		43.97476653684007	±	2.5660556692661527		13.686083846304543	±	1.9692421763823178		0.19874923253356436	±	0.14054262912707596		10.648929627760829	±	1.2610417692161926		2.8384049860101506	±	0.7250489634103734		385.2983552173646	±	11.11947099385128
	lhy/cca1	0.38839844244417515	±	0.011305696234666993		87.66875134320924	±	0.6616859381627912		0.03501709356264063	±	0.242174820606473	40.89549360576745	±	1.9435358548963813		44.33442846772069	±	3.3974269961757013		11.9428502143466	±	0.672672873341385		0.36242565386416775	±	0.1765679870409028		9.58158791724833	±	0.3098373581141258		1.9988366432341014	±	0.602751078749119		634.1374553492403	±	43.20750489431593	*
























	28 Days
		EtOH				Water				Neutral			Acid				Basic				Insol				Starch				Protein				Cell wall				Assimilation
	Col-0	0.5880259875091256	±	0.6459486258134546		84.42303230327809	±	0.9454792960122854		-0.011283180088240322	±	0.04600689857442257	35.31368052143015	±	1.2292815130884744		52.56335314113098	±	1.7231085974105698		14.988941709212778	±	1.534435552054548		2.3975149934639166	±	1.1625119763112792		6.961297655899314	±	1.2587212175737352		5.6301290598495495	±	0.623644796595388		236.18149035759	±	44.99250796500788
	prr7/9	0.3899983303242155	±	0.04070830504918652		80.2234016014161	±	0.931996207270137	*	-0.049914728317449544	±	0.043263813419269194	42.36684284922242	±	0.49936209900898565	*	39.84839696862185	±	1.5357704875892044	*	19.386600068259693	±	0.9175596921584043	*	1.76407086626919	±	0.7713042077211923		11.4853564462326	±	0.5844917804360515		6.137172755757903	±	1.2115580541968216		251.1481780493945	±	9.341755122604958
	WS	0.2536554662649522	±	0.05355653265583025		79.21534714686528	±	0.6420048242744627		0.03445720354619943	±	0.006916216453753687	27.845114794391446	±	0.6582689932241103		55.287673698952375	±	0.5755836605401101		20.530997386869757	±	0.6388519927509423		2.7089373257498015	±	0.46141917231777185		10.09663130794166	±	0.8587341525976818		7.725428753178295	±	0.6580109495003585		187.0311434726941	±	8.504953696810352
	lhy/cca1 (fl)	0.37449445032369405	±	0.0476570285585143	*	90.39347475944122	±	0.7999185055915877	*	-0.0644640485532166	±	0.08221556939998122	42.27574574274346	±	1.5321959297539127	*	55.66874642094534	±	1.345439531202377		9.232030790235092	±	0.8463401810763305	*	0.9244950273178872	±	0.1393066545719478	*	5.30059632860077	±	0.5442363496880182	*	3.006939434316434	±	0.6140001306069902	*	259.6764283147239	±	19.6934717521723	*



















Col
prr7prr9







EtOH soluble
Water soluble
Water insoluble







Neutral
Acidic
Basic







Starch
Protein
Cell wall





Experiment number 106a		16.03.15

	DAP	plants labelled	Pools	Plants per pool	n
Col-0	18	12	3	4	3
WS	18	12	3	4	3
lhy/cca1	18	12	3	4	3
prr7/9	18	12	3	4	3

	Time	ZT
Pulse start	1899-12-31T4:15:0+0000	"19:15"
Pulse end	1899-12-31T5:15:0+0000	"20:15"

Label	600 ul	150 uC

EtOH volume	3 ml

Experiment number 106a		16.03.15

	DAP	plants labelled	Pools	Plants per pool	n
Col-0	28	4	4	1	4
WS	28	4	4	1	4
lhy/cca1	28	4	4	1	4
prr7/9	28	4	4	1	4

	Time	ZT
Pulse start	1899-12-31T5:50:0+0000	1899-12-31T20:50:0+0000
Pulse end	1899-12-31T6:50:0+0000	1899-12-31T21:50:0+0000

Label	600 ul	150 uC

EtOH volume	5 ml



Experiment 106a

DAP	Genotype	Sample number	Mass (g)	Mass (mg)	Notes
Experiment 106a
18	Col-0	1	0.067	67	Harvested Col-0 and prr7/9 first alternating between
18	Col-0	2	0.043	43
18	Col-0	3	0.0757	75.7
18	WS	1	0.0334	33.4	Then WS and lhy so effective 10 min chase
18	WS	2	0.0266	26.599999999999998
18	WS	3	0.0297	29.7
18	lhy/cca1	1	0.0351	35.1
18	lhy/cca1	2	0.0249	24.9
18	lhy/cca1	3	0.0293	29.3
18	prr7/9	1	0.0366	36.6
18	prr7/9	2	0.0274	27.400000000000002
18	prr7/9	3	0.0244	24.400000000000002


Experiment 106b
28	Col-0	1	0.3097	309.7	This harvest was much faster due to less plants
28	Col-0	2	0.3156	315.59999999999997	Col-0 prr first again then WS lhy
28	Col-0	3	0.2043	204.3
28	Col-0	4	0.2577	257.7
28	WS	1	0.2673	267.29999999999995
28	WS	2	0.4418	441.8
28	WS	3	0.252	252
28	WS	4	0.275	275	lost some sample during grinding… will probably have to eliminate from calculations
28	lhy/cca1	1	0.0965	96.5
28	lhy/cca1	2	0.1125	112.5
28	lhy/cca1	3	0.0864	86.4
28	lhy/cca1	4	0.0912	91.2
28	prr7/9	1	0.14	140
28	prr7/9	2	0.1475	147.5
28	prr7/9	3	0.1356	135.6
28	prr7/9	4	0.1603	160.3

100


Fill in all rawdata into one table!

Scintillation counting data			Experiment name	Date

					lsc	calculation		total counts					Water sol. Dowex					Insoluble digests																																														Satndard deviations
no.	sp.	probe name	fraction	solubilized in	dpm	calc dpm		sample	total counts	Assimilation	EtOH	Water	Neutral	Acid	Basic	recovery water sol %	Insol	Starch	Protein	remaining	total counts		%										WT
		all blanks						name	sum of sol, insol, EtOH sol of leaf and rosette		leaf										sum of sol, insol, EtOH sol of leaf		EtOH	Water	Neutral	Acid	Basic	Insol	Starch	Protein	Remaining			EtOH			Water			Neutral			Acid			Basic			Insol			Starch			Protein			Cell wall			Assimilation			EtOH	Water	Neutral	Acid	Basic	Insol	Starch	Protein	Cell wall	Assimilation
		sol blank			12.05		A.t.	Col-0 1	23422.6	349.59104477611936	116.93999999999998	20546.66	15	9790.4	10151.4	97.1291684390553	2759	50	2083.6000000000004	625.3999999999996	sum of sol, insol, EtOH sol of rosette	Col-0 1	0.49926139711219075	87.72151682562995	0.06404071281582745	41.798946316805136	43.34019280523938	11.779221777257863	0.21346904271942485	8.895681948203874	2.670070786334564		Col-0	0.44783639721781787	±	0.04543542414093131	86.59258816279475	±	0.8032073167483322	0.08771570416643622	±	0.07048602777232704	40.52926432957474	±	2.5262081546687742	43.74125700701987	±	3.5226742845625796	12.959575439987427	±	0.8369099758029841	0.9177178972887473	±	0.7978715645510204	8.785055784169613	±	0.7612789559370693	3.256801758529068	±	0.4211181412095605	377.7537984343283	±	26.55853220216979	0.04543542414093131	0.8032073167483322	0.07048602777232704	2.5262081546687742	3.5226742845625796	0.8369099758029841	0.7978715645510204	0.7612789559370693	0.4211181412095605	26.55853220216979
		EtOH blank			16.85			Col-0 2	17774.34	413.3567441860465	80.96	15271.58	32.59999999999998	7605	7046	96.14984173215869	2421.7999999999997	90	1716.4999999999998	615.3		Col-0 2	0.4554880800074714	85.91925213538167	0.18341046699905583	42.78639881987179	39.64141565875301	13.625259784610847	0.5063479150280685	9.657179957174217	3.461731912408562		Ws	0.44192273868188675	±	0.03291544537103846	85.87199341501356	±	1.9861423039490287	-0.046192116560937246	±	0.10293146401224013	42.88782457399689	±	2.168316353512296	43.97476653684007	±	2.5660556692661527	13.686083846304543	±	1.9692421763823178	0.19874923253356436	±	0.14054262912707596	10.648929627760829	±	1.2610417692161926	2.8384049860101506	±	0.7250489634103734	385.2983552173646	±	11.11947099385128	0.03291544537103846	1.9861423039490287	0.10293146401224013	2.168316353512296	2.5660556692661527	1.9692421763823178	0.14054262912707596	1.2610417692161926	0.7250489634103734	11.11947099385128
		NCS blank			14			Col-0 3	28032.739999999998	370.313606340819	108.98	24146.559999999998	4.399999999999977	10372.8	13523.600000000002	98.98221527207191	3777.2	570	2187.2	1020		Col-0 3	0.3887597145337916	86.13699552737263	0.015695932684425346	37.00244785204728	48.24216255706722	13.474244758093572	2.0333367341187487	7.802305447130747	3.6386025768440766		lhy/cca1	0.38839844244417515	±	0.011305696234666993	87.66875134320924	±	0.6616859381627912	0.03501709356264063	±	0.242174820606473	40.89549360576745	±	1.9435358548963813	44.33442846772069	±	3.3974269961757013	11.9428502143466	±	0.672672873341385	0.36242565386416775	±	0.1765679870409028	9.58158791724833	±	0.3098373581141258	1.9988366432341014	±	0.602751078749119	634.1374553492403	±	43.20750489431593	0.011305696234666993	0.6616859381627912	0.242174820606473	1.9435358548963813	3.3974269961757013	0.672672873341385	0.1765679870409028	0.3098373581141258	0.602751078749119	43.20750489431593
		acidic			9.04			WS 1	12644.41	378.5751497005988	56.16	11152.3	11.199999999999974	5671.4	6019	104.92544138877182	1435.95	37.5	1138.6499999999999	259.8000000000002		WS 1	0.44414883731229843	88.19944940096059	0.08857669120188269	44.853022007353445	47.60206288786903	11.35640176172712	0.2965737428634472	9.005165128305709	2.0546628905579634		prr7/9	0.7886351792247885	±	0.4024416923388231	82.5043008714346	±	0.8754554070053538	0.19421013151321795	±	0.13739603188905347	44.00025207583705	±	0.5584576389951075	34.65610012126357	±	1.4587549861856766	16.707063949340608	±	1.2604142979692203	0.03431775909206191	±	0.1617962073106456	13.719813498010012	±	1.0717337605792359	2.9529326922385333	±	0.42253706955437775	275.6563134032681	±	41.8160822643151	0.4024416923388231	0.8754554070053538	0.13739603188905347	0.5584576389951075	1.4587549861856766	1.2604142979692203	0.1617962073106456	1.0717337605792359	0.42253706955437775	41.8160822643151
		basic			9.56			WS 2	10010.88	376.3488721804511	48.16	8343.72	-6.600000000000001	3991	4211.2	98.2247726433773	1619	30	1208.3000000000002	380.6999999999998		WS 2	0.4810765886715254	83.34651898734178	-0.0659282700421941	39.86662511187828	42.06623193964966	16.172404423986702	0.2996739547372459	12.06986798363381	3.802862485615649
		Neutral			12.13			WS 3	11908.84	400.9710437710438	47.7	10249.939999999999	-19.200000000000017	5233.199999999999	5032.200000000001	99.96351198153357	1611.2	0	1294.7	316.5		WS 3	0.40054279006183646	86.07001185673835	-0.16122477084250034	43.94382660275895	42.256004783001536	13.52944535319981	0	10.87175577134297	2.657689581856839
1	A.t.	Col-0 1	EtOH Soluble	EtOH	75.32	116.93999999999998		lhy/cca1 1	23187.48	660.6119658119658	88.94	20215.34	0.5999999999999872	8846.2	10546.000000000002	95.93110974141422	2883.2	100	2301	482.1999999999998		lhy/cca1 1	0.38356906399488	87.18213449671978	0.0025876033100620988	38.15076066911972	45.48144084652581	12.434296439285337	0.43126721834369236	9.923458694088362	2.079570526853284
2		Col-0 2	EtOH Soluble	EtOH	57.33	80.96		lhy/cca1 2	16647.98	668.5935742971888	67.25999999999999	14750.82	-40.40000000000003	7015.799999999999	7958	101.23776169731582	1829.9	20	1606.2	203.70000000000005		lhy/cca1 2	0.40401297935244995	88.60426309978749	-0.2426720839405143	42.14204966608561	47.80159514848048	10.99172392086007	0.1201346950200565	9.648017357060738	1.2235718687792756
3		Col-0 3	EtOH Soluble	EtOH	71.34	108.98	L.j.	lhy/cca1 3	16794.96	573.2068259385665	63.42	14648.539999999999	58.4	7120	6671	94.5445757734218	2083	90	1540.65	452.3499999999999		lhy/cca1 3	0.3776132839851956	87.21985643312041	0.34772336462843617	42.393670482097015	39.72024940815578	12.402530282894393	0.5358750482287544	9.173287700595894	2.6933675340697443
4		WS 1	EtOH Soluble	EtOH	44.93	56.16		prr7/9 1	8084.26	220.88142076502731	109.46	6759	0	3526	2929.2000000000003	95.50525225625094	1215.8000000000002	20	1004.9000000000001	190.9000000000001		prr7/9 1	1.3539891097020629	83.60691021812757	0	43.61561849816805	36.23337200931193	15.039100672170367	0.24739431933163952	12.430327574818229	2.3613787780205002
5		WS 2	EtOH Soluble	EtOH	40.93	48.16		prr7/9 2	8832.34	322.3481751824817	49.7	7281.4400000000005	26.199999999999974	3956	3093	97.16759322331846	1501.2	0	1207.8	293.4000000000001		prr7/9 2	0.5627047871798413	82.44066691273207	0.2966371312698557	44.78994241616604	35.0190323289185	16.996628300088084	0	13.67474531098214	3.321882989105946
6		WS 3	EtOH Soluble	EtOH	40.7	47.7		prr7/9 3	6923.240000000001	283.7393442622951	31.099999999999994	5640.04	19.79999999999997	3018.2	2265	94.02415585705066	1252.1000000000001	-10	1042.25	219.85000000000014		prr7/9 3	0.44921164079246123	81.46532548344416	0.28599326326979807	43.59519531317706	32.71589602556028	18.085462875763373	-0.1444410420554538	15.054367608229672	3.175536309589153
7		lhy/cca1 1	EtOH Soluble	EtOH	61.32	88.94
8		lhy/cca1 2	EtOH Soluble	EtOH	50.48	67.25999999999999
9		lhy/cca1 3	EtOH Soluble	EtOH	48.56	63.42
10		prr7/9 1	EtOH Soluble	EtOH	71.58	109.46
		prr7/9 2	EtOH Soluble	EtOH	41.7	49.7
		prr7/9 3	EtOH Soluble	EtOH	32.4	31.099999999999994
		Col-0 1	Water Soluble	Water	10290.18	20546.66
		Col-0 2	Water Soluble	Water	7652.64	15271.58
		Col-0 3	Water Soluble	Water	12090.13	24146.559999999998
		WS 1	Water Soluble	Water	5593	11152.3
		WS 2	Water Soluble	Water	4188.71	8343.72																Two-tailed, two-sample t-test with equal variance
		WS 3	Water Soluble	Water	5141.82	10249.939999999999
		lhy/cca1 1	Water Soluble	Water	10124.52	20215.34																ttest	EtOH	Water	Neutral	Acid	Basic	Insol	Starch	Protein	Remaining	Assimilation
		lhy/cca1 2	Water Soluble	Water	7392.26	14750.82																Prr7/9 vs Col0	0.2998353803969166	0.008241201060973332	0.3846402903638842	0.13064376218695486	0.028048331743674746	0.024831108838473667	0.19967655775644202	0.006051467594094865	0.5111513428475137	0.043472001427598425
		lhy/cca1 3	Water Soluble	Water	7341.12	14648.539999999999																Lhy/cca1 vs WS	0.09528064520715347	0.2915970498763503	0.6819462655631556	0.3880273420485383	0.9106658436775091	0.3017182753092314	0.36300821464075933	0.3096937760174065	0.2764066541673317	0.0013969928804853035
		prr7/9 1	Water Soluble	Water	3396.35	6759
		prr7/9 2	Water Soluble	Water	3657.57	7281.4400000000005
		prr7/9 3	Water Soluble	Water	2836.87	5640.04
		Col-0 1	Insoluble	NCS	292.75	2759
		Col-0 2	Insoluble	NCS	259.03	2421.7999999999997
		Col-0 3	Insoluble	NCS	394.57	3777.2
		WS 1	Insoluble	NCS	208.31	1435.95
		WS 2	Insoluble	NCS	178.75	1619
		WS 3	Insoluble	NCS	177.97	1611.2
		lhy/cca1 1	Insoluble	NCS	305.17	2883.2
		lhy/cca1 2	Insoluble	NCS	199.84	1829.9
		lhy/cca1 3	Insoluble	NCS	225.15	2083
		prr7/9 1	Insoluble	NCS	138.43	1215.8000000000002
		prr7/9 2	Insoluble	NCS	166.97	1501.2
		prr7/9 3	Insoluble	NCS	142.06	1252.1000000000001
		Col-0 1	Starch		5	50
		Col-0 2	Starch		9	90
		Col-0 3	Starch		57	570
		WS 1	Starch		5	37.5
		WS 2	Starch		3	30
		WS 3	Starch		0	0
		lhy/cca1 1	Starch		10	100
		lhy/cca1 2	Starch		2	20
		lhy/cca1 3	Starch		9	90
		prr7/9 1	Starch		2	20
		prr7/9 2	Starch		0	0
		prr7/9 3	Starch		-1	-10
		Col-0 1	prot		416.72	2083.6000000000004
		Col-0 2	prot		343.29999999999995	1716.4999999999998
		Col-0 3	prot		437.44	2187.2
		WS 1	prot		151.82	1138.6499999999999
		WS 2	prot		241.66000000000003	1208.3000000000002
		WS 3	prot		258.94	1294.7
		lhy/cca1 1	prot		460.20000000000005	2301
		lhy/cca1 2	prot		321.24	1606.2
		lhy/cca1 3	prot		308.13	1540.65
		prr7/9 1	prot		200.98000000000002	1004.9000000000001
		prr7/9 2	prot		241.56	1207.8
		prr7/9 3	prot		208.45000000000002	1042.25
		Col-0 1	Neutral		12.88	15
		Col-0 2	Neutral		13.76	32.59999999999998
		Col-0 3	Neutral		12.35	4.399999999999977
		WS 1	Neutral		12.69	11.199999999999974
		WS 2	Neutral		11.8	-6.600000000000001
		WS 3	Neutral		11.17	-19.200000000000017
		lhy/cca1 1	Neutral		12.16	0.5999999999999872
		lhy/cca1 2	Neutral		10.11	-40.40000000000003
		lhy/cca1 3	Neutral		15.05	58.4
		prr7/9 1	Neutral		12.13	0
		prr7/9 2	Neutral		13.44	26.199999999999974
		prr7/9 3	Neutral		13.12	19.79999999999997
		Col-0 1	Basic		517.13	10151.4
		Col-0 2	Basic		361.86	7046
		Col-0 3	Basic		685.74	13523.600000000002
		WS 1	Basic		310.51	6019
		WS 2	Basic		220.12	4211.2
		WS 3	Basic		261.17	5032.200000000001
		lhy/cca1 1	Basic		536.86	10546.000000000002
		lhy/cca1 2	Basic		407.46	7958
		lhy/cca1 3	Basic		343.11	6671
		prr7/9 1	Basic		156.02	2929.2000000000003
		prr7/9 2	Basic		164.21	3093
		prr7/9 3	Basic		122.81	2265
		Col-0 1	Acidic		498.56	9790.4
		Col-0 2	Acidic		389.29	7605

100


Fill in all rawdata into one table!

Scintillation counting data			Experiment name	Date

					lsc	calculation		total counts					Water sol. Dowex					Insoluble digests
no.	sp.	probe name	fraction	solubilized in	dpm	calc dpm		sample	total counts	Assimilation	EtOH	Water	Neutral	Acid	Basic	recovery water sol %	Insol	Starch	Protein	remaining	total counts		%										WT																															Satndard deviations
		all blanks						name	sum of sol, insol, EtOH sol of leaf and rosette		leaf										sum of sol, insol, EtOH sol of leaf		EtOH	Water	Neutral	Acid	Basic	Insol	Starch	Protein	Remaining			EtOH			Water			Neutral			Acid			Basic			Insol			Starch			Protein			Cell wall			Assimilation
		sol blank			10.72		A.t.	Col-0 1	52870.28	170.7144979011947	137.64000000000001	44728.64	8.79999999999999	17781.600000000002	28506.199999999997	103.50549446618543	8004	820	4712.549999999999	2471.4500000000007	sum of sol, insol, EtOH sol of rosette	Col-0 1	0.2603352961247794	84.60072464151884	0.016644511812685673	33.63250582368772	53.91724802667963	15.138940062356394	1.5509658734548029	8.91341978896272	4.67455439993887		Col-0	0.5880259875091256	±	0.6459486258134546	84.42303230327809	±	0.9454792960122854	-0.011283180088240322	±	0.04600689857442257	35.31368052143015	±	1.2292815130884744	52.56335314113098	±	1.7231085974105698	14.988941709212778	±	1.534435552054548	2.3975149934639166	±	1.1625119763112792	6.961297655899314	±	1.2587212175737352	5.6301290598495495	±	0.623644796595388	236.18149035759	±	44.99250796500788	EtOH	Water	Neutral	Acid	Base	Insol				Assimilation
		EtOH blank			15.33			Col-0 2	71754.51999999999	227.359062103929	142.70000000000002	59680.72	35	24917.4	37041.2	103.8754224144749	11931.099999999999	2890	5064.45	3976.6499999999987		Col-0 2	0.19887248914772204	83.17346419431139	0.048777414997689346	34.72589601324071	51.62211384035459	16.627663316540893	4.027620838380635	7.0580222681442235	5.5420202100160365		Ws	0.2536554662649522	±	0.05355653265583025	79.21534714686528	±	0.6420048242744627	0.03445720354619943	±	0.006916216453753687	27.845114794391446	±	0.6582689932241103	55.287673698952375	±	0.5755836605401101	20.530997386869757	±	0.6388519927509423	2.7089373257498015	±	0.46141917231777185	10.09663130794166	±	0.8587341525976818	7.725428753178295	±	0.6580109495003585	187.0311434726941	±	8.504953696810352	0.6459486258134546	0.9454792960122854	0.04600689857442257	1.2292815130884744	1.7231085974105698	1.534435552054548	1.1625119763112792	1.2587212175737352	0.623644796595388	44.99250796500788
		NCS blank			14			Col-0 3	60349.240000000005	295.3952031326481	1029.4399999999998	51778.5	-27.200000000000024	21763	32882.4	105.48432264356826	7541.3	650	3306.8000000000006	3584.4999999999995		Col-0 3	1.7058044144383586	85.79809787165505	-0.04507099012348792	36.06176316387745	54.486850207227135	12.496097713906588	1.077064102215703	5.479439343395211	5.939594268295672		lhy/cca1 (fl)	0.37449445032369405	±	0.0476570285585143	90.39347475944122	±	0.7999185055915877	-0.0644640485532166	±	0.08221556939998122	42.27574574274346	±	1.5321959297539127	55.66874642094534	±	1.345439531202377	9.232030790235092	±	0.8463401810763305	0.9244950273178872	±	0.1393066545719478	5.30059632860077	±	0.5442363496880182	3.006939434316434	±	0.6140001306069902	259.6764283147239	±	19.6934717521723	0.05355653265583025	0.6420048242744627	0.006916216453753687	0.6582689932241103	0.5755836605401101	0.6388519927509423	0.46141917231777185	0.8587341525976818	0.6580109495003585	8.504953696810352
		acidic			9.04			Col-0 4	64748.979999999996	251.25719829258827	121.14000000000001	54466.74	-42.40000000000002	23850	32521.600000000002	103.41944460050298	10161.099999999999	1900	4140.25	4120.8499999999985		Col-0 4	0.18709175032564224	84.11984250562712	-0.06548365703984839	36.8345570849147	50.22720049026256	15.69306574404724	2.934409159804525	6.394309223095098	6.364347361147618		prr7/9	0.3899983303242155	±	0.04070830504918652	80.2234016014161	±	0.931996207270137	-0.049914728317449544	±	0.043263813419269194	42.36684284922242	±	0.49936209900898565	39.84839696862185	±	1.5357704875892044	19.386600068259693	±	0.9175596921584043	1.76407086626919	±	0.7713042077211923	11.4853564462326	±	0.5844917804360515	6.137172755757903	±	1.2115580541968216	251.1481780493945	±	9.341755122604958	0.0476570285585143	0.7999185055915877	0.08221556939998122	1.5321959297539127	1.345439531202377	0.8463401810763305	0.1393066545719478	0.5442363496880182	0.6140001306069902	19.6934717521723
		basic			9.56			WS 1	48059.079999999994	179.79453797231577	135.5	37651.079999999994	13.199999999999967	13763.000000000002	26491.199999999997	106.9488577751289	10272.5	1070	5430.9	3771.6000000000004		WS 1	0.281944639805839	78.34332242731237	0.02746619369326248	28.637668469725185	55.12215381567853	21.37473293288178	2.226426306953858	11.300466009752997	7.847840616174927																																	0.04070830504918652	0.931996207270137	0.043263813419269194	0.49936209900898565	1.5357704875892044	0.9175596921584043	0.7713042077211923	0.5844917804360515	1.2115580541968216	9.341755122604958
		nucleotide			12.13			WS 2	80553.07999999999	182.3292892711634	143.92000000000002	63985.259999999995	25.799999999999983	21770.2	44046.6	102.90276229244049	16423.9	2070	7536.599999999999	6817.300000000002		WS 2	0.1786648008989849	79.43241897144095	0.03202857047800033	27.025906396130356	54.680218310708916	20.38891622766008	2.5697341430023535	9.356066831957264	8.46311525270046
1	A.t.	Col-0 1	EtOH Soluble	EtOH	84.15	137.64000000000001		WS 3	50140.34	198.96960317460315	150.6	40047.24	21.999999999999993	13975	28109	105.14082868132735	9942.5	1670	4830.200000000001	3442.2999999999993		WS 3	0.3003569580900329	79.87030004184255	0.04387684646733547	27.87176951731879	56.060648970469686	19.82934300006741	3.3306515272931936	9.633361082114721	6.865330390659496
2		Col-0 2	EtOH Soluble	EtOH	86.68	142.70000000000002		WS 4														WS4 (dropped)
3		Col-0 3	EtOH Soluble	EtOH	530.05	1029.4399999999998	L.j.	lhy/cca1 1	26879.3	278.5419689119171	118.62	24573.18	-25.80000000000002	10864.8	15517.4	107.256773441614	2187.5	230	1306.3	651.2		lhy/cca1 1	0.4413061352044138	91.42046109831729	-0.09598464245720692	40.420695479420964	57.72992600253727	8.138232766478295	0.8556770451611464	4.859873583017415	2.4226821382997326
4		Col-0 4	EtOH Soluble	EtOH	75.9	121.14000000000001		lhy/cca1 2	26616.660000000003	236.59253333333336	105.70000000000002	24171.06	-1.8000000000000327	10956.000000000002	14901.000000000002	106.96758851287449	2339.9	310	1262.5500000000002	767.3499999999999		lhy/cca1 2	0.3971196987150153	90.81176977126356	-0.006762681718893477	41.16218939566422	55.98373349623882	8.791110530021422	1.1646840738094109	4.743457668993781	2.88296878721823
5		WS 1	EtOH Soluble	EtOH	83.08	135.5		lhy/cca1 3	21056.3	243.7071759259259	67.88000000000001	18801.82	-38.60000000000003	9296.2	11432.2	110.04147470829952	2186.6	180	1157.6000000000001	848.9999999999998		lhy/cca1 3	0.32237382636075673	89.2930856798203	-0.18331805682859778	44.14925699196915	54.29348935947912	10.384540493818951	0.8548510422058956	5.49764203587525	4.032047415737807
6		WS 2	EtOH Soluble	EtOH	87.29	143.92000000000002		lhy/cca1 4	25523.600000000006	279.86403508771934	86.06	22983.640000000003	7.199999999999989	11069.8	13953.2	108.9044207096874	2453.8999999999996	210	1557.3	686.5999999999997		lhy/cca1 4	0.33717814101459037	90.0485824883637	0.02820918679183182	43.3708411039195	54.66783682552617	9.614239370621695	0.8227679480950961	6.101412026516634	2.6900593960099655
7		WS 3	EtOH Soluble	EtOH	90.63	150.6		prr7/9 1	34218.16	244.4154285714286	143.54	26916.420000000002	-34.20000000000002	14325.4	14375.800000000001	106.50376238742001	7158.200000000001	190	4170.299999999999	2797.9000000000015		prr7/9 1	0.41948485833253446	78.66121381161348	-0.09994692876531062	41.86490448346725	42.012194694279295	20.919301330053983	0.5552607153628365	12.187388217250719	8.176652397440428
8		WS 4	EtOH Soluble	EtOH	127.26	223.86		prr7/9 2	38888.3	263.64949152542374	159.32	31538.780000000002	-1.6000000000000014	16586.600000000002	14977.2	100.07425778676284	7190.2	670	4233.25	2286.95		prr7/9 2	0.40968620381965776	81.10094810007124	-0.004114348017269979	42.65190301453137	38.513383202659924	18.48936569610911	1.7228832322318022	10.885664840067577	5.880817623809731
9		lhy/cca1 1	EtOH Soluble	EtOH	74.64	118.62		prr7/9 3	34761.28	256.35162241887906	142.88	27963.4	-29.800000000000004	14568.2	13309.400000000001	99.58660248753729	6655	740	4150	1765		prr7/9 3	0.41103204484990197	80.44410332415838	-0.0857275681447864	41.90927376667373	38.28800320356443	19.144864630991727	2.1288053834611387	11.9385707315726	5.0774885159579854
10		lhy/cca1 2	EtOH Soluble	EtOH	68.18	105.70000000000002		prr7/9 4	38500.24	240.17616968184652	123.12	31064.82	-3.8000000000000256	16571	15623.400000000001	103.62397078109579	7312.3	1020	4208	2084.3		prr7/9 4	0.31979021429476806	80.68734116982128	-0.009870068342431178	43.041290132217355	40.580006773983754	18.992868615883953	2.649334134020983	10.929801996039505	5.4137324858234654
		lhy/cca1 3	EtOH Soluble	EtOH	49.27	67.88000000000001
		lhy/cca1 4	EtOH Soluble	EtOH	58.36	86.06
		prr7/9 1			87.1	143.54
		prr7/9 2			94.99	159.32
		prr7/9 3			86.77	142.88
		prr7/9 4			76.89	123.12
		Col-0 1	Water Soluble	Water	22375.04	44728.64
		Col-0 2	Water Soluble	Water	29851.08	59680.72
		Col-0 3	Water Soluble	Water	25899.97	51778.5
		Col-0 4	Water Soluble	Water	27244.09	54466.74																Two-tailed, two-sample t-test with equal variance
		WS 1	Water Soluble	Water	18836.26	37651.079999999994
		WS 2	Water Soluble	Water	32003.35	63985.259999999995
		WS 3	Water Soluble	Water	20034.34	40047.24																ttest	EtOH	Water	Neutral	Acid	Basic	Insol	Starch	Protein	Remaining	Assimilation
		WS 4	Water Soluble	Water	20359.67	40697.899999999994																Prr7/9 vs Col0	0.6151810269907949	0.0015448574064050302	0.33014592141268195	9.25792527660067E-5	7.565642537897797E-5	0.005318868364509797	0.46153825164228	0.0013235065995955335	0.5430903846148476	0.5931088790468318
		lhy/cca1 1	Water Soluble	Water	12297.31	24573.18																Lhy/cca1 vs WS	0.044886523160916764	1.3670533582610238E-5	0.13932525035394902	4.946086735724792E-5	0.7134396626372725	1.557524748602247E-5	0.0016255109370621064	6.194479313541211E-4	4.276757917820235E-4	0.00390767901324832
		lhy/cca1 2	Water Soluble	Water	12096.25	24171.06
		lhy/cca1 3	Water Soluble	Water	9411.63	18801.82
		lhy/cca1 4	Water Soluble	Water	11502.54	22983.640000000003
		prr7/9 1	Water Soluble		13468.93	26916.420000000002
		prr7/9 2	Water Soluble		15780.11	31538.780000000002
		prr7/9 3	Water Soluble		13992.42	27963.4
		prr7/9 4	Water Soluble		15543.13	31064.82
		Col-0 1	Insoluble	NCS	814.4	8004
		Col-0 2	Insoluble	NCS	1207.11	11931.099999999999
		Col-0 3	Insoluble	NCS	768.13	7541.3
		Col-0 4	Insoluble	NCS	1030.11	10161.099999999999
		WS 1	Insoluble	NCS	1041.25	10272.5
		WS 2	Insoluble	NCS	1656.39	16423.9
		WS 3	Insoluble	NCS	1008.25	9942.5
		WS 4	Insoluble	NCS	926.08	9120.800000000001
		lhy/cca1 1	Insoluble	NCS	232.75	2187.5
		lhy/cca1 2	Insoluble	NCS	247.99	2339.9
		lhy/cca1 3	Insoluble	NCS	232.66	2186.6
		lhy/cca1 4	Insoluble	NCS	259.39	2453.8999999999996
		prr7/9 1	Insoluble		729.82	7158.200000000001
		prr7/9 2	Insoluble		733.02	7190.2
		prr7/9 3	Insoluble		679.5	6655
		prr7/9 4	Insoluble		745.23	7312.3
		Col-0 1	Starch		82	820
		Col-0 2	Starch		289	2890
		Col-0 3	Starch		65	650
		Col-0 4	Starch		190	1900
		WS 1	Starch		107	1070
		WS 2	Starch		207	2070
		WS 3	Starch		167	1670
		WS 4	Starch		111	1110
		lhy/cca1 1	Starch		23	230
		lhy/cca1 2	Starch		31	310
		lhy/cca1 3	Starch		18	180
		lhy/cca1 4	Starch		21	210
		prr7/9 1	Starch		19	190
		prr7/9 2	Starch		67	670
		prr7/9 3	Starch		74	740
		prr7/9 4	Starch		102	1020
		Col-0 1	prot		942.5099999999999	4712.549999999999
		Col-0 2	prot		1012.89	5064.45
		Col-0 3	prot		661.3600000000001	3306.8000000000006
		Col-0 4	prot		828.05	4140.25
		WS 1	prot		1086.1799999999998	5430.9
		WS 2	prot		1507.32	7536.599999999999
		WS 3	prot		966.0400000000001	4830.200000000001
		WS 4	prot		977.4799999999999	4887.4
		lhy/cca1 1	prot		261.26	1306.3
		lhy/cca1 2	prot		252.51000000000002	1262.5500000000002
		lhy/cca1 3	prot		231.52	1157.6000000000001
		lhy/cca1 4	prot		311.46	1557.3
		prr7/9 1	prot		834.06	4170.299999999999
		prr7/9 2	prot		846.6500000000001	4233.25
		prr7/9 3	prot		830	4150
		prr7/9 4	prot		841.6	4208
		Col-0 1	Neutral		12.57	8.79999999999999
		Col-0 2	Neutral		13.88	35
		Col-0 3	Neutral		10.77	-27.200000000000024
		Col-0 4	Neutral		10.01	-42.40000000000002
		WS 1	Neutral		12.79	13.199999999999967
		WS 2	Neutral		13.42	25.799999999999983