Molecular analysis of bunch architecture in grapevine Eva Zyprian a - PowerPoint PPT Presentation

Molecular analysis of bunch architecture in grapevine Eva Zyprian a , Robert Richter a , Susanne Rossmann b , Klaus Theres b , Reinhard Töpfer a a Julius Kühn-Institut, Institute for Grapevine Breeding Geilweilerhof, 76833 Siebeldingen, Germany b Max Planck Institute, 50829 Cologne, Germany GBG 2018, Bordeaux, France, July 15-20, 2018

Why study bunch architecture? Loose clusters • Dry off quickly • Don´t squeeze each other • Have less micro cracks • Have intact wax layers • Are better covered by protective sprays • Are more resilient to Botrytis bunch rot and other infections Represent a physical factor of • resistance Zyprian et al., GBG 2018, Bordeaux, France, July 15-20, 2018

QTL Analysis in population GF.GA- 47-42 x `Villard blanc´ • 150 F 1 individuals with considerable (transgressive) segregation of cluster architecture traits • genetic map available (Zyprian et al., 2016) • eight vines per genotype • 46 females, 104 hermaphrodite plants OIV class 9 OIV class 1 Individual 0324 Individual 0212 Zyprian et al., GBG 2018, Bordeaux, France, July 15-20, 2018

QTL Analysis in population GF.GA-47-42 x `Villard blanc´ Sampling: 6 basally inserted clusters at 6 different trunks BBCH81 – BBCH87 Zyprian et al., GBG 2018, Bordeaux, France, July 15-20, 2018

Phenotyping of cluster architecture sub-traits, over two to four years Peduncle • Image based assessment of sub-traits using Wing  BAT software for volume (Kicherer et al. 2013)  ImageJ for length parameters Internodes • Reference measurements: volume, weight Pedicel Berry •  in total 19 ampelometric Rachis measurements Zyprian et al., GBG 2018, Bordeaux, France, July 15-20, 2018 Zyprian et al., GBG 2018, Bordeaux, France, July 15-20, 2018

Cluster architecture sub-traits Notation Sub-trait [Unit] 2013 n=12 2014 n=3 2015 n=6 2016 n=6 2017 n=6 berry number per bunch BN x x x x Brix sugar content of juice [°Bx] x x BW berry weight [g] x x BW/RW berry weight/rachis weight x x cluster weight [g] CW x x x x mean single berry volume [cm³] MBV x x x x OIV204 compactness x x x x pedicel length [cm] PED x x x x PL peduncle length [cm] x x x x L1I length 1st internode of rachis [cm] x x x L2I length 2nd internode of rachis [cm] x x x L3I length 3rd internode of rachis [cm] x x RD diameter 2nd internode of rachis [cm] x x rachis length [cm] RL x x x x RW rachis weight x x shoulder length SL x x x TBV total berry volume x x x x TLL total length of laterals x x presence/ absence of a “shoulder” Wing x x Sub-traits with large effect on CA (OIV204) after statistical evaluation and modeling Zyprian et al., GBG 2018, Bordeaux, France, July 15-20, 2018

Statistical evaluation of cluster architecture sub-traits Flower sex affects cluster architecture: Females are more loose! Zyprian et al., GBG 2018, Bordeaux, France, July 15-20, 2018

23 QTLs for large effect sub-traits in eight clusters 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 rachis pedicel density berry Zyprian et al., GBG 2018, Bordeaux, France, July 15-20, 2018

Fanizza et al. 2005 table grape Cabezas et al. 2006 table grape Costantini et al. 2008 table grape Marguerit et al. 2009 hybrid cross newly identified QTL region Doligez et al. 2013 table/wine Markers flanking QTL clusters are Correa et al. 2014 table grape available Zyprian et al., GBG 2018, Bordeaux, France, July 15-20, 2018

Expression Analysis in `Pinot noir´ French P.noir gene Origin pool Trait compact mixed berries loose INRA / Frank / Geisenheim / Freiburg / M- /L-type Geisenheim / Group France Baden Hessen Baden Wädenswil Hessen ENTAV Frank Frank Gm-20- PN WE- Fr12-L Gm-1- Clones Fr1801 777 Charisma Classic 13 40024 171 / 13-L 86 Foto: Antes Foto: E. Wennesheimer Worms Foto: R.Richter , JKI Geilweilerhof Zyprian et al., GBG 2018, Bordeaux, France, July 15-20, 2018

Plants in three different locations Hessen Hessen Antes Heppenheim 1995 Pfalz 125AA /1,8m² Mannheim JKI Siebeldingen 2005 125AA/1,8 m² Sibbus Jechtingen Baden 1997 125AA/2,2m² Freiburg Sampling harmonized according to degree/days (Molitor, 2014) Pfalz Baden Zyprian et al., GBG 2018, Bordeaux, France, July 15-20, 2018 Foto: google earth

Phenotyping in ‘Pinot noir’ • Eight of 20 sub-traits show significant differences between clones • Grouping in loose and compact clustered clones reveals four sub- traits as significantly different Variance analysis (ANOVA) indicates • Berry volume • Pedicel length • Rachis length • Total berry volume as determinants of CA Zyprian et al., GBG 2018, Bordeaux, France, July 15-20, 2018

Investigation of rachis elongation Weekly rachis length 2015 Weekly rachis length 2016 Difference to week before 2015 Difference to week before 2016 Measurement of rachis length at Geilweilerhof each week from May to August 2015 and 2016 at the same bunch (n = 16) compact MB loose compact MB loose

RNA-Seq, RT-qPCR: Candidate gene expression in ‘Pinot noir’ clones 2015/ 2016/ 2017 A candidate gene is Hessen consistently higher Pfalz Mannheim expressed in loose PN Fold change of VIT_16s0039g01450 GRF6 expression clones over three years and 2016 BBCH 71 Fold-change of GRF 6 expression in PN clones in ‘Pinot Noir’ clones at BBCH 71 located at three Baden Freiburg three locations field Trials fold-change (compared toFrank Charisma) three bio repeats (endogeneous control: GAPDH) (endogenous control: GAPDH) bars indicate SEM 8,00 5,00 Fold change (compared to Frank Charisma) 4,50 7,00 2016 2015 4,00 6,00 Fold change Fold change 3,50 5,00 3,00 4,00 2,50 2,00 3,00 1,50 2,00 1,00 1,00 0,50 0,00 Entav_777-… Frank Classic-… Frank Classic-… Gm-20-13-… Freiburg-1801-… 0,00 Entav_777-Baden Gm-20-13-Pfalz Gm-20-13-Baden Gm-1-86-Hessen Gm-1-86-Pfalz WE-171-Pfalz compact MB loose compact MB loose Zyprian et al., GBG 2018, Bordeaux, France, July 15-20, 2018

Conclusions • Eight QTL regions contain genetic determinants of cluster architecture in GF.GA.47-42 x `Villard blanc´ Markers from these regions may now be tested for transferability of their linkage to the trait in extended breeding material • Two candidate genes are differentially expressed in loose versus compact `Pinot noir´ clones, over three locations Zyprian et al., GBG 2018, Bordeaux, France, July 15-20, 2018

Thank you for your attention! Thanks to… Grapevine nursery Antes, Heppenheim Grapevine nursery Sibbus, Jechtingen Doreen Gabriel Sarina Elser Florian Rist Margit Schneider Daniel Zendler Anna Schwandner Anna Kicherer