Breeding new flood-tolerant rice varieties
description
Transcript of Breeding new flood-tolerant rice varieties
![Page 1: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/1.jpg)
Bert Collard
Rice Seminar Series
Current position: Scientist II (breeder)Since March 2011
Education and trainingPhD in Agricultural Science (1998 – 2002 ) - University of Melbourne, VIC,
AustraliaBachelor of Science (Honours) (1993 – 1997) - University of Melbourne, VIC,
Australia
Work experienceDurum wheat breeder (2009-2011) - Department of Primary Industries NSWResearch Scientist (2008) - Department of Primary Industries VICResearch Scientist (2006-2007) - Queensland Department of Primary IndustriesPostdoctoral Research Fellow (2005-2006) - PBGB, IRRIPostdoctoral Research Fellow (2003-2004) - University of Southern QueenslandResearch Assistant (2000-2002) - RMIT University, VIC
![Page 2: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/2.jpg)
Research highlights• Senior durum wheat breeder/ program leader for Australian Durum wheat
breeding program• >10 years experience in molecular genetics research for disease
resistance, abiotic stress tolerances and quality traits in bread wheat, chickpea, barley, sorghum and rice
• Experience in DNA marker validation research within Australian Wheat and Barley Molecular Marker Program
• Working with Dave Mackill at IRRI (2005-2006) in marker-assisted backcrossing program for Sub1 and abiotic stress tolerances
IRRI claim to fame:• Rice breeding course co-ordinator• Highest water bill by research staff in PBGB (or even IRRI ?)
![Page 3: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/3.jpg)
Breeding new flood tolerant rice varieties
Bert CollardIRRI
![Page 4: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/4.jpg)
PART I: Past achievements
4
PART II: Current activities
PART III: Future directions
![Page 5: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/5.jpg)
Flooding
5
![Page 6: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/6.jpg)
Types of flooding1. Crop establishment (anaerobic germination)
2. Flash-flooding (short duration ~2 weeks)
3. Medium deep water (Stagnant, slow rise)
6
![Page 7: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/7.jpg)
FR13A
7
![Page 8: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/8.jpg)
Sub1 is major QTL for submergence tolerance
0 10 20 30 40
LOD score
50cM
100cM
150cM
OPN4
OPAB16
C1232
RZ698
OPS14RG553
R1016RZ206
RZ422
C985
RG570
RG451
RZ404
Sub-1(t)
1200
850
900
OPH7950
OPQ1600
~ 69% of the variation explained by a single QTL on chromosome 9: Sub1
Xu KN & Mackill DJ (1996). Mol. Breeding 2:
219-224.
8
![Page 9: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/9.jpg)
9
Xu et al. (2006) Nature 442: 705-708
![Page 10: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/10.jpg)
“Mega-varieties” of rice• extremely popular varieties which have higher yields
and farmers prefer• Problem: susceptible to abiotic stresses• Solution: use backcrossing strategy
BR11 (Bangladesh)
Swarna (India) IR64 (Asia)
![Page 11: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/11.jpg)
“…there are many undesirable aspects associated with precipitous changes in varieties. Farmers are reluctant to rapidly shift their production from an old proven variety to an unknown new one. Their reluctance is based on their familiarity with the old variety which permits them to exploit to the maximum potential yielding ability. They know the best rates and dates of seeding of the old variety for their local conditions. ...
...The conventional backcross method .... [produces] new varieties which are phenotypically similar to the recurrent parent and is thereby readily received by both farmer and miller.”
11
Normal Borlaug 1957
![Page 12: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/12.jpg)
DNA markers improve efficiency of backcrossing
12
![Page 13: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/13.jpg)
CONVENTIONAL BACKCROSSINGx P2P1
DONORVARIETY
P1 x F1
P1 x BC1
P1 x BC2
P1 x BC3
P1 x BC4
P1 x BC5
P1 x BC6
BC6F2
• Select target gene. Discard ~50% BC1
• Visually select BC1 progeny that closely resemble variety
~6
YE
AR
S
Repeat process until at least BC6
13
![Page 14: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/14.jpg)
MAS
MARKER ASSISTED BACKCROSSINGx P2P1
DONORVARIETY
P1 x F1
P1 x BC1
P1 x BC2
P1 x BC3
BC3Fn fixed lines
3 -
4 Y
EA
RS MAS
MAS
MAS
14
![Page 15: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/15.jpg)
1 2 3 4 5 6 7 8 9 10 11 12
SUB1
CONVENTIONAL BACKCROSSING
Unwanted donor chromosome segments
15
![Page 16: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/16.jpg)
1 2 3 4 5 6 7 8 9 10 11 12
SUB1
MARKER ASSISTED BACKCROSSING
High level of precision
16
![Page 17: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/17.jpg)
C25 straight after de-submergence 2011 WS
17
![Page 18: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/18.jpg)
Swarna-Sub1 after submergence stress
1 week 2 weeks 3 weeks
18
![Page 19: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/19.jpg)
Submergence tolerance screening
Swarna Swarna Sub1 19
![Page 20: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/20.jpg)
Short video
20
![Page 21: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/21.jpg)
Rigorous evaluation of Sub1 varieties lines for:• Submergence tolerance• Yield and agronomic traits• Grain quality
Rigorous evaluation of Sub1 varieties lines for:• Submergence tolerance• Yield and agronomic traits• Grain quality
Name Year developed Country
Swarna-Sub1 2006India, Bangladesh, Nepal
IR64-Sub1 2007 All Asia
Samba Mahsuri-Sub1 2007
India, Bangladesh, Nepal
TDK 1-Sub1 2007 LaosBR11-Sub1 2007 BangladeshCR1009-Sub1 2007 India
Ciherang-Sub1 2009 Indonesia, Bangladesh
PSBRc18-Sub1 2009 Philippines
21
![Page 22: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/22.jpg)
22
![Page 24: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/24.jpg)
The price of “Sub1 fame”• Frequent seed requests and visitors!
24Allan Salabsabin
INGER
![Page 25: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/25.jpg)
Looking ahead
Promotion• Replace Swarna (approx. 5 M
ha area) with Swarna-Sub1• Take Swarna-Sub1 to
additional flood prone areas where Swarna cannot be cultivated
Tracking of diffusion of Swarna-Sub1 using
• GIS & Remote sensing• Seed production &
diffusion through formal & informal systems
• Ground survey
25
Umesh SinghIRRI, STRASA
South Asia Project Co-ordinator
![Page 26: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/26.jpg)
IRRI Submergence group
![Page 27: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/27.jpg)
27
BR11-Sub1“Pavel” Iftekharaudaula
Ciherang-Sub1Nurul Hidayatun
PSB Rc18-Sub1Darlene Sanchez
Eloi Estoy Freddie
Varoy Septi
![Page 28: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/28.jpg)
PART II:Current activities
28
![Page 29: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/29.jpg)
Sub1 variety enhancements in progress
29
Country Variety Stage Target date
Nepal Sabitri-Sub1 BC2F2/ BC1F3
2014
Philippines*(*PhilRice collaboration)
PSBRc82-Sub1 BC2F1/ BC1F2
2014-2015
Pakistan IR6-Sub1Super Basmati-Sub1
BC2F1 2014-2015
![Page 30: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/30.jpg)
New SNP genotyping platforms
Illumina “BeadXpress”
384 SNPs
“Fluidigm EP”24, 48 or 96 SNPs with
flexible assays30
![Page 31: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/31.jpg)
Improving disease resistanceBacterial leaf blight
Marker assisted pyramiding of 5 genes:
SUB1 + Xa4 + xa5 + xa13 + Xa21
606)
31
Collaboration with Dr. Nollie Vera-Cruz
![Page 32: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/32.jpg)
Developing Swarna-Sub1 with photoperiod sensitivity
• May overcome problems due to delays in flowering
• Used MABC for se1 (chr. 6)
• Field observations 2012 dry and wet season
32
![Page 33: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/33.jpg)
33
Screening of Sub1 in elite IRRI material
![Page 34: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/34.jpg)
Expanded submergence screening facilities at IRRI
34
![Page 35: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/35.jpg)
Stagnant flooding
• Water depth: 50-60 cm for up to several months
• Yield losses: ~<60% • Common in South
(Eastern India, Bangladesh, Nepal) and SE Asia (Thailand, Cambodia) and parts of Africa
35
![Page 36: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/36.jpg)
Sub1 has NO effect on stagnant flooding tolerance
• Sub1 has no affect on tolerance
Swarna-Sub1
36
“Sacobia” (Philippines, 1997); “Schwe Pyi Tan” (Myanmar, 2005)
“Popoul” (Cambodia 1999)
Indonesia (2008)
• 3 varieties with Sub1 + stagnant flooding tolerance:
![Page 37: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/37.jpg)
Limited knowledge on stagnant flooding tolerance
• Correlation with plant height = 0.17 (NS)
• Correlation with elongation ability = 0.69 (P<0.001)
37
Dr. Yoichi Kato (CESD)
Dr. S.R. Das (OUAT)
• Breeder’s eyes:– Tillering R2 = 0.12– Maturity R2 = 0.39
![Page 38: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/38.jpg)
Promising elite lines with Sub1 + stagnant flooding tolerance
38IRRI 154(NSIC Rc222)
![Page 39: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/39.jpg)
F2
F3
F4
F5
F6
OBSERVATION YIELD TRIAL (OYT)
ADVANCED YIELD TRIALS (AYT)
<10,000 PLANTS
>200,000 PLANTSn = 1,000/ pop. x 50 crosses/ season
~400
80
~8,000 lines
3000-4000 lines
1000 lines
MAS
Quality
SUBMERGENCE
SUBMERGENCE
BACTERIAL BLIGHT
Yield, SUBMERGENCE, Blast, BPH, GLH
Forward breeding
Yield under NORMAL + STAGNANT FLOODING
BACTERIAL BLIGHT
Selection for intermediate height + yield + yield components
39Yield under NORMAL
+ STAGNANT FLOODING
![Page 40: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/40.jpg)
Promising material
• Selection for yield under SF stress
• Developed SUB1 + SFT lines outperforming both Sub1 varieties and checks
![Page 41: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/41.jpg)
International collaboration is critical for developing new varieties
IRRI CORE BREEDING PROGRAM
INGER Flood prone nurseries
OVERSEAS PARTNERSObservational or replicated
yield trials
BEST ELITE LINES
IRRI multi-environment
testing (MET)
n = 10 – 30 per season
NEW VARIETIES
Eastern Indian Rainfed Lowland Shuttle breeding network (CRRI)
BRRI41
![Page 42: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/42.jpg)
South Asian Breeding Network
BRRI
NEPAL
EIRLSBN
BRRI
![Page 43: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/43.jpg)
PART III:Future directions
43
![Page 44: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/44.jpg)
Enhancing submergence tolerance
West Bengal 2011 WS
Several regions were affected by complete
submergence for 20 to 26 days
44
![Page 45: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/45.jpg)
Current level of tolerance
Swarna Swarna Sub1 45
![Page 46: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/46.jpg)
New QTLs
PSB Rc18(IRRI 105)
PSB Rc18 Sub146
#1
![Page 47: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/47.jpg)
Stagnant flooding
Research needs:1. Set of elite breeding lines2. Phenotyping methods3. Germplasm evaluation4. Physiological trait
characterization5. Molecular genetic
information
47
#2
![Page 48: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/48.jpg)
Stagnant flooding
48
#2
![Page 49: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/49.jpg)
49
![Page 50: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/50.jpg)
Germplasm screening
50
#3New sources of submergence tolerance
Elite germplasm for stagnant flooding tolerance
![Page 51: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/51.jpg)
Improved routine phenotyping
SES
#4
![Page 52: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/52.jpg)
Precision phenotyping#4
![Page 53: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/53.jpg)
Submergence screening
53
![Page 54: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/54.jpg)
54
![Page 55: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/55.jpg)
DAY 1
DAY 7
DAY 10
DAY 14
DAY 15
DAY 16
DAY 17
DAY 18
DAY 12
SAMPLE #1
SAMPLE #2
SAMPLE #3
55
Submergence computer model
Input:• Water temp.• pH• Dissolved O2• EC• Turbidity
OUTPUT #1:Drain water now.85% survival tolerant check
OUTPUT #2:10% yield loss for Swarna-Sub1
![Page 56: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/56.jpg)
56
Li et al. (2010) Field Crops Research 118: 221-227.
16.8% 40.7%
Precision phenotyping - digital image analysis
![Page 57: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/57.jpg)
Multiple stress tolerances
Submergence + salinity tolerance
57
Submergence + drought tolerance
IRRI 119 IR10F388
#5
![Page 58: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/58.jpg)
Priorities and constraints
Dr. J.N. Reddy, Central Rice Research Institute (CRRI), Cuttack, Odisha
58
![Page 59: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/59.jpg)
Towards a rainfed breeding program
Disease & insect resistance
Salinity
Drought
Flood tolerance
SUB1 automatically built into all new varieties
59
![Page 60: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/60.jpg)
60
Eero Nissila, Head PBGB
![Page 61: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/61.jpg)
61
MOLECULAR BREEDING
PHYSIOLOGY + AGRONOMY
FLOOD TOLERANCE TRAIT DEVELOPMENT TEAM
ABIOTIC STRESS SCREENING $
![Page 62: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/62.jpg)
Summary
1. Sub1 varieties have had a big impact in a short time
2. Currently developing new Sub1 varieties with multiple types of flooding tolerance
3. “Building” new rainfed varieties with multiple stress tolerances next priority
4. Sub1 will be a “default” gene for all breeding programs
62
![Page 63: Breeding new flood-tolerant rice varieties](https://reader030.fdocuments.in/reader030/viewer/2022020122/544e68bcb1af9fe65a8b48d3/html5/thumbnails/63.jpg)
63
• Marlyn Rala
SUB Breeding team• Jerome Carandang• Julius “Jojo” Borgonia• Richard Daif• Richard Formaran
Lab Team• Jean Gonzaga• Eloi Suiton• Darlene Sanchez• Kashif Aslam Physiology (CESD)• Yoichi Kato• Ella Evangelista• Abdel Ismail
ES• Leigh Vial• Caling Balingbing
PBGB• Varoy Pamplona• Dr. S.R. Das• Endang Septiningsih• Glenn Gregorio• Allan Sallabsabin• Rafiq Islam
MMAL • Jade Dilla-Ermita• Mike Thomson
Pathology• Nollie Vera-Cruz• “Abe” Ona• Ruby Burgos
• Umesh Singh• Dave Mackill
Thank you!