Celi, L.

Ferrandino, A.

Lovisolo, Claudio

Patono, DAVIDE LUCIEN

Said Pullicino, D.

English

Institutional Research Information System University of Turin

18 October 2023AperTO - Archivio Istituzionale Open Access dell'Università di TorinoOriginal Citation:Design and set up of a plant growth chamber for stable isotope labeling to investigate carbonattraction toward fruit sinks and plant reserves upon and after drought stressTerms of use:Open Access(Article begins on next page)Anyone can freely access the full text of works made available as "Open Access". Works made availableunder a Creative Commons license can be used according to the terms and conditions of said license. Useof all other works requires consent of the right holder (author or publisher) if not exempted from copyrightprotection by the applicable law.Availability:This is a pre print version of the following article:This version is available http://hdl.handle.net/2318/1679986 since 2018-10-31T15:30:21ZDesign	and	set	up	of	a	plant	growth	chamber	 for	stable	 isotope	 labeling	 to	 investigate	carbon	attraction	toward	fruit	sinks	and	plant	reserves	upon	and	after	drought	stress.		D.	L.	Patono1,	D.	Said	Pullicino1,	A.	Firbus1,	A.	Ferrandino1,	G.	Gambino2,	L.	Celi1,	Claudio	Lovisolo1,2	1Dept.	Agricultural,	Forest	and	Food	Sciences,	University	of	Turin,	Grugliasco,	Italy	2	Institute	for	Sustainable	Plant	Protection,	National	Research	Council,	Turin,	Italy	Results	of	preliminary	investigation	for	setting	air	fluxes	into	the	growth	chamber	for	stable	isotope	labeling.																											Photosynthetic	 assimilation,	 stomatal	 regulation	 and	 respiration	 are	 checked	 in	 the	 various	phases	to	size	13CO2	enrichment	flows	in	the	chamber.	Maximum	assimilation	ranges	from	6	to	13	μmol	m-2	 s-1,	 reduced	possibly	by	 stomatal	 control	 3	o	 4	 times	 at	 the	end	of	 the	drought	period.	Light	responses	are	evaluated	to	optimizing	chamber	illumination.	A	model	optimizing	light	and	temperature	is	possible	for	plants	in	the	various	stages	of	the	experiment.	In	the	same	sinks	taken	for	isotopic	analysis,	the	expression	of	genes	involved	in	carbohydrate	transport	will	be	investigated.	Genes	encoding	proteins	that	regulate	the	delivery	of	sucrose	to	the	sinks	and	which	catalyze	the	hydrolysis	of	the	sucrose	discharged	to	trigger	respiration	or	C	storage	will	be	analyzed.	Financial	support:	Cassa	di	Risparmio	di	Torino	Foundation.	Open	questions:	-	How	are	neosynthesized	sugars	translocated	at	different	rates	depending	on	the	hydration	status	of	the	plant?	-	 Are	 these	 sugars	 during	 stress	 periods	 redistributed	 following	 different	 ratios	 toward	different	sinks?	-	 Is	 quality	 of	 the	 berry	 related	 to	 variations	 in	 delivery	 dynamics	 of	 sugars	 during	 water	stress?	-	 Is	 rate	 of	 respiration	 of	 sugars	 different	 depending	 on	 whether	 or	 not	 water	 stress	 is	present?	Experimental	set	up	Three	 treatments	x	 three	 replicates	 (irrigated	controls	 IRR,	drought	 stressed	plants	WS,	and	re-hydrated	recovering	plants	REC)	simultaneously	in	the	13C	labelling	chamber	(=	9	plants	in	the	chamber).Three	plants	with	optimum	hydration	act	as	a	control	for	both	the	water	stress	line	and	the	recovery	line.		All	plants	during	the	translocation	time	(period	following	the		pulse		of		13C)		are		all	 	exposed		to	 	 the	 	 same	 environmental	 conditions,	 minimizing	 environmental	 (solar	 radiation,	temperature,	air	humidity)	influence	on	C	translocation	rates.	At	the	end	of	the	13C	pulse:	leaf	sampling	to	know	the	amount	of	C13	assimilated	by	the	square	meter	of	leaf	surface.	Time	 kinetics:	 Sampling	 leaves,	 berries,	 roots	 and	 soil	 at	 subsequent	 time	 points	 to	understand	deliveries	of	C	toward	sinks.	Once	a	maximum	of	13C	has	reached	the	various	districts,	by		repeating	sampling	it	is	possible	an	estimation	of	13C	leaving	the	district	(respiration	/re-translocation).	Grapevine	is	a	plant	that	adapts	to	periods	of	drought,	which	are	correlated	with	berry	quality.		In	perennial	plants,	the	root	orchestrates	the	defense	adaptations	to	drought,	acting	as	a	sink	of	the	carbon	(C)	allocated	during	growth	slowdowns,	C	which	can	floematically	be	released	in	the	post-drought	periods.	In	crops	oriented	to	fruit	production	(and	in	this	case	in	the	grapevine)	the	root	sink	competes	with	fruits	in	receiving	photosynthates	during	the	growing	season,	and	the	competition	increases	with	water	stress.	In	addition,	 the	current	climatic	variations	 lead	 to	 the	alternation	of	extreme	events	with	periods	of	drought.	The	atmospheric	CO2	concentration,	 the	 triggering	factor	of	the	greenhouse	effect,	is	constantly	rising,	and	the	C	cycle	in	plants	could	partly	mitigate	the	effect,	but	the	high	temperatures	affect	both	photorespiration	and	C	dynamics	in	plant	and	soil.			We	aim	to	study	C	allocation	kinetics	in	grapevine	organs	in	a	controlled	water	deficit	system	basing	on	studies	of	C	allocation,	trough	pulse-chasing	isotopic	strategy.	The	isotope	acts	as	a	tracer	of	the	floematic	flows	that	are	oriented	towards	different	sinks	during	drought/rehydration	cycles.	Grapevine	fits	well	the	subject	of	investigation,	especially	in	relation	to	two	well-known	and	deeply	studied	factors:	i)  root:	grapevines	(Vitis	vinifera	L.	cultivars)	in	commercial	vineyards	are	grafted	on	different	rootstocks,	belonging	to	pure	or	hybrid	genotypes	of	the	genus	Vitis.	This	confers	to	root	systems	a	wide	spectrum	of	vigour,	water	relations,	and	strategies	of	resistance/resilience	to	water	stress.		ii)  fruit:	grape	berries	belong	to	hundreds	of	cultivars,	spread	all	over	the	world.	Both	fresh	consume	(table	grapes)	and	berry	to	win	transformation	(oenological	viticulture)	point	to	maximize	fruit	quality,	driving	in-field	ripening	dynamics	toward	accumulation	onto	berries	(skin,	pulp	and	seeds)	of	sugars	and	secondary	metabolites.	This	to	improve	taste	and	flavor	of	either	fresh	berries	or	their	derived	wines,	and	to	protect	them	from	detrimental	reactions	happening	during	fruit	conservation,	oenological	technologies,	and	wine	ageing.	105033003780365 300 365 485N.B.: misure in mm, salvo dove diversamente indicato440010002310350110060010001330 1330312456Vista in pianta, livello piante con indicazione configurazione a pieno caricoVista lateraleSTRUTTURA PRINCIPALE LABELLING CHAMBER - TELAIO STRUTTURA PRINCIPALE LABELLING CHAMBER - SUPERFICIVista lateraleVista in pianta, livello piante con indicazione delle superfici di appoggio7LEGENDA21 DIREZIONE E UFFICI:  VIA TIRRENO, 151 – 10136  TORINO TEL 011.323.913   FAX 011.323.991 E-MAIL  info@inteksrl.net  –  WEB  www.inteksrl.netPART.  IVA  06346640011Sede Legale: Via San Quintino, 31 - 10121 TORINO - Cap. Soc. € 10.400,00 i.v. - CCIAA R.D. 779600 - Iscriz. Trib. di Torino n. 2718/ 92   ISO 9001 – Cert. N. SC 13-3461    ISO 14001    OHSAS 18001    UNI EN 1090 – UNI 11339    Reg. (CE) 303/2008 e 304/2008    UNI 11352 105033003780365 300 365 485N.B.: misure in mm, salvo dove diversamente indicato440010002310350110060010001330 1330312456Vista in pianta, livello piante con indicazione configurazione a pieno caricoVista lateraleSTRUTTURA PRINCIPALE LABELLING CHAMBER - TELAIO STRUTTURA PRINCIPALE LABELLING CHAMBER - SUPERFICIVista lateraleVista in pianta, livello piante con indicazione delle superfici di appoggio7LEGENDA21 DIREZIONE E UFFICI:  VIA TIRRENO, 151 – 10136  TORINO TEL 011.323.913   FAX 011.323.991 E-MAIL  info@inteksrl.net  –  WEB  www.inteksrl.netPART.  IVA  06346640011Sede Legale: Via San Quintino, 31 - 10121 TORINO - Cap. Soc. € 10.400,00 i.v. - CCIAA R.D. 779600 - Iscriz. Trib. di Torino n. 2718/ 92   ISO 9001 – Cert. N. SC 13-3461    ISO 14001    OHSAS 18001    UNI EN 1090 – UNI 11339    Reg. (CE) 303/2008 e 304/2008    UNI 11352 Lateral	view	Plan	view	105033003780365 300 365 485N.B.: misure in mm, salvo dove diversamente indicato440010002310350110060010001330 1330312456Vista in pianta, livello piante con indicazione configurazione a pieno caricoVista lateraleSTRUTTURA PRINCIPALE LABELLING CHAMBER - TELAIO STRUTTURA PRINCIPALE LABELLING CHAMBER - SUPERFICIVista lateraleVista in pianta, livello piante con indicazione delle superfici di appoggio7LEGENDA21 DIREZIONE E UFFICI:  VIA TIRRENO, 151 – 10136  TORINO TEL 011.323.913   FAX 011.323.991 E-MAIL  info@inteksrl.net  –  WEB  www.inteksrl.netPART.  IVA  06346640011Sede Legale: Via San Quintino, 31 - 10121 TORINO - Cap. Soc. € 10.400,00 i.v. - CCIAA R.D. 779600 - Iscriz. Trib. di Torino n. 2718/ 92   ISO 9001 – Cert. N. SC 13-3461    ISO 14001    OHSAS 18001    UNI EN 1090 – UNI 11339    Reg. (CE) 303/2008 e 304/2008    UNI 11352 Design	of	the	13C	labelling	chamber	[1]	Fan	coil	unit	Sabiana	type	mod.	Futura	FSC,	tot	coil	power	2,15	kW.	[2]	Refrigeration	unit	Carrier	type	mod.	30	AWH	006,	tot	coil	power		4,73	kW.	[3]	Polycarbonate	panels	anti-UV	treated,	thickness	6mm.	[4]	Deflector	made	of	polycarbonate	anti-UV	treated,	thickness	6	mm.	System	design	and	installation	made	with	INTEK	Innovazioni	Tecnologiche	srl.	0	200	400	600	800	1000	3	 8	 13	 18	 23	PPFD (𝛍mol*m2s-1) hour	of	the	day	Natural	and	artificial	modulation	of	PPFD	inside	labelling	chamber	y	=	0.6382x	+	0.2496	0	20	40	60	80	100	120	140	160	180	200	0	 100	 200	 300	 400	Area	(cm2)	Square	leaf	diameter	(cm2)	0	0.1	0.2	0.3	0.4	0.5	Area	(m2)	Total	area	of	the	groups	of	classes	∅	1-5	cm	 ∅	6-10	cm	 ∅	11-15	cm	0	2	4	6	8	10	12	14	0	 400	 800	 1200	 1600	 2000	 2400	A (𝛍mol*m2s-1 ) PPFD	(𝛍mol*m2s-1 )	Net	photosynthesis	at	different	light	intensity	∅	11-15	cm	 ∅	6-10	cm	 ∅	1-5	cm	0	5	10	15	20	25	30	1	 2	 3	 4	 5	 6	 7	 8	 9	 10	 11	 12	 13	 14	 15	Absolute	average	frequency	of	classes	Leaf	diameter	classes	(cm)	Absolute	average	frequency	of	diameter	classes	per	plant	Pulse	duration	 Total	assimilated	modeling	1234567(1)	The	longest	diameter	of	every	leaf	per	plant	was	measured	and	reported	in	this	graph.	The	dotted	lines	represent	following	grouping	of	diameter.	(2)	Regression	area/square	diameter	for	leaves	of	grapevine	cv	Barbera.	(3)	By	combining	absolute	average	frequencies	of	diameter	classes	with	relative	leaf	area	it	is	possible	to	calculate	total	leaf	area	of	the	plant.	(4)	The	total	area	of	the	three	groups	of	diameter	were	counted	separately	as	representing	different	leaf	development	stages,	corresponding	to	different	photosynthetic	activity.	(5)	Ambient	PPFD	can	be	tuned	artificially	with	LED	lamps	to	allow	appropriate	light	intensity	inside	the	chamber.	(6)	Net	photosynthesis	at	different	light	intensity	of	the	three	groups	were	measured	to	allow	prediction	of	photosynthetic	rate	inside	the	chamber.	(7)	By	combining	total	leaf	area	of	grops	with	light	intinsity	trend	of	the	chamber	and	photosynthetic	rates	of	groups	is	possible	to	predict	total	CO2	assimilation	per	plant,	in	order	to	design	experimental	pulses.	

Design and set up of a plant growth chamber for stable isotope labeling to investigate carbon attraction toward fruit sinks and plant reserves upon and after drought stress

https://iris.unito.it/bitstream/2318/1679986/1/poster%20Patono%20et%20al%2018%20FISV%20Roma%20Sapienza.pdf

Design and set up of a plant growth chamber for stable isotope labeling to investigate carbon attraction toward fruit sinks and plant reserves upon and after drought stress

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