DUS Test Guidelines for Maize

DUS, PPV & FR Act 2001

1. DUS Testing Guidelines for Maize Ph.D . (Genetics & Plant Breeding) Reg. No. : 04-00899-2011 Course Code: GP 691 Presented by: K. Satish

2. CONTENTS Maize Introduction Production constraints Importance of maize PPV & FR Act, 2001 Objectives Criteria for protection Benefits DUS Testing Assessment of DUS Characteristics and Symbols Technical questionnaire Table of characteristics of maize Conclusions Future thrust

3. Introduction 1 Taxonomically, Zea mays L . belongs to tribe Maydeae , family Poaceae and genus Zea ( Doebley et al., 2004 ). The center of origin of Zea mays L. ( 2n=20 ) is reported to be Mexico and Central America (Chopra et al., 2002) The main maize growing countries include U.S.A., China, Brazil, European Union, Ukraine, Argentina, India and Mexico. Maize is the third most important cereal crop in India after rice and wheat. It accounts for 9 per cent of total food grain production in the country. Nine states viz . Karnataka, Andhra Pradesh, Tamil Nadu, Rajasthan, Maharashtra, Bihar, Uttar Pradesh, Madhya Pradesh and Gujarat account for 85 per cent of India’s maize production and 80 per cent of area under cultivation ( FAO, 2013 )

4. Table: 1 Comparison of Area, Production and Yield of Maize, 2013-14 Source: Directorate of Economics and Statistics, Department of Agriculture and Cooperation, GOI (2013-14) USDA (2013-14) State Area under cultivation Million Hectare Production Million tonnes Yield Kg / Hectare World 177.4 960.2 5508 India 9.43 23.35 2583 Gujarat 0.46 0.69 1501 2

5. Maize is a monoecious plant. Male inflorescence called tassel Female inflorescence known as silk Generally protandrous Each floret contains three anthers and each plant produces about 3-5 million pollen grains . Wind borne nature of the pollen and protandry leads to cross pollination but, there may also be reported 5 % self pollination. Plant Morphology of Maize Chopra et al., 2002 3

6. Importance of Maize Top ranking cereal in terms of production and productivity globally Highest potential of carbohydrate production per unit/day ( 62.2 %) Great significance as human food, animal feed and industrial products Grown in unmatched diversity of environments Queen of cereals 4

7. Maize Utilization for Various Purposes Source of human food (24%), animal feed (11%), poultry feed (52%), starch (11%), brewery (1%) and seed (1%) In addition more than 500 industrial products like starch, oil, protein, alcohol, acetic acid, sucrose, glucose, paper, rayon, dyes, synthetic rubber, resin etc. are being manufactured from it. 5 Ranum et al., 2014

8. PPV & FR Act, 2001 Government of India enacted the Plant variety protection & Farmer’s Rights Act on Oct. 30 th 2001 Plant variety protection under PPV & FR Act, provides legal protection to the plant variety Article 27.3(b) – Member country to provide protection either by a patent or Sui generis system or in combination To a breeder in the form of Plant Breeder’s Rights To a farmers in the form of Farmer’s Rights To a Researcher in the form of Researcher’s Rights PPV & FR Act, 2001 was enacted by Indian Government based on UPOV Convention Act. PPV & FR Authority, India 6

9. Conti… Objectives Registrable plant varieties New Variety Extant Variety Farmer’s Variety Essentially Derived Varieties (EDV) Criteria for protection Novelty – [(Article 15.3(a)] variety was not sold out before 1 year of registration Distinctness - [(Article 15.3(b)] at least one essential character Uniformity - [(Article 15.3(c)] sufficiently uniform Stablilty - [(Article 15.3(d)] not susceptible to change after repeated propagation Who can apply? Benefits 7

10. DUS Test Why? Essential requirement for registration of new variety(s) under PPV & FR Act, 2001 Variety be examined for compliance with Distinctiveness, Uniformity and Stability criteria needed for registration. Sujay et al ., 2001 8 How? Involves comparison of new (candidate) variety against existing varieties for recording a number of morphological/physiological characters, by growing new and existing varieties side-by-side . What is DUS Test? Plant variety protection can only be granted in respect of a new plant variety after examination, for that UPOV Convention Acts laid down criteria viz. novelty (N), distinctness (D), uniformity (U) and stablity (S) ‘ DUS’ or ‘NDUS’ in short.

11. Must be clearly distinguishable for at least one essential character from the varieties of common knowledge in India and out side. Essential character is a heritable trait which is determined by one or more genes or other heritable determinants that contribute to the principal features, performance or value of the plant variety. DISTINCTNESS 9

12. Uniform, when subjected to the variation that may be expected and allowed due to the specific nature of the reproduction of the crop such as vegetative, self- and cross-pollinated. UNIFORMITY STABILITY Stable when all essential characters remain unchanged after repeated propagation or a specified cycle of propagation. 10

13. Guidelines for Conduct of Test for Distinctiveness, Uniformity and Stability On Maize UPOV Species Code: ZEAAA_MAY ( Zea mays L.) Protection of Plant Varieties and Farmer’s Rights Authority (PPV & FRA) Department of Agriculture & Co-operation Ministry of Agriculture, Government of India 11

14. Requirements for DUS Testing Planting material Candidate Variety Reference Collection Staff Scientific/Technical/Secretarial/Field Facilities Land/Gardens(flowers)/Orchard(trees) DUS Test Guidelines DUS test centres Laboratory with equipments RHS colour Chart Data base of Extant/Notified varieties ( NORV and IINDUS ) Software for database / analysis ( OOS, PLOD_PLAN, ANALIST 3.1, DUSTNT ) 12 source: www.plantauthority.gov.in

15. Rice Wheat Maize Sorghum Pearlmillet DRR, Hyderabad DWR, Karnal DMR, New Delhi ( Kharif ) NRCS, Hyderabad MPKV, Rahuri CRRI, Cuttack IARI, Indore ANGRAU, Hyderabad ( Kharif and Rabi ) MPKV, Rahuri Mandore IARI, Karnal CSUAT, Kanpur RAU, Dholi ( Kharif and Rabi ) GBPUA&T, Pantnagar AAU, Jorhat UAS, Dharwad VPKAS, Almora , Uttarakhand (Hill region Kharif ) TNAU, Coimbatore PAU, Ludhiana DUS testing centre in India for cereals source: www.plantauthority.gov.in Table: 2 DUS test centres 13

16. Planting material for DUS testing Crop Quantity (g) Maize 3000(varieties & hybrids) 1500 (parental lines) Wheat 3000(varieties & hybrid) Rice 3000 (varieties & hybrids) 1500 (parental lines) Sorghum 1500 (varieties & hybrids) 500 (parental lines) Pearl millet 1000 (varieties & hybrids) 500 (parental lines) Annonymous , 2007 A. Germination capacity i Inbred line & single cross hybrids 80% (minimum) ii Varieties & double cross hybrids 90% (minimum) B. Moisture content 10-12% (maximum) C. Physical purity 98% (minimum) Table - 3: Seed Quantity for Test Table - 4: Seed Standards, Moisture and Physical purity 14

17. Table - 5: Test plot design for Maize ● Number of rows i ) Inbred line & single cross hybrids 4 ii) Varieties & other hybrids 8 ● Row length 6 m ● Row to row distance 75 cm ● Plant to plant distance 20 cm ● Number of replication 3 Number of growing cycle Test location Condition favoring normal growth and expression . Observations should not be recorded on plants in border rows . Additional test for special purpose may be established. New Delhi PPV & FR Authority Conduct of Test 15

18. Assessment of DUS Distinctness Distinctness is established by comparing the candidate variety with existing varieties/reference collections. Consistent differences Clear differences If similar state of expression - variety is not distinct Uniformity Population standard of 3 % and an acceptance probability of at least 95 % should be applied. In sample size of 100 plants, 3 off-types are allowed. Same should applied to clear case of out-crossed plants in inbred lines as well as single cross hybrids. For visually assessed characteristics, the uniformity of the candidate variety is checked against known comparable varieties. UPOV , 2009 16

19. Conti… Stability Require 2-3 years to perform a test of stability which leads to the same certainty as the testing of the distinctness and homogeneity. Submitted sample to be homogenous during the test , the material can also be considered to be stable. In case of doubt inbred and open pollinated varieties may tested by growing a further generation , or by testing a new seed stock . All observation on ear should be made on the upper well-developed ear. Latest Royal Horticultural Society (RHS) colour chart should be used. 17 Selvi et al., 2013

20. 1. Reference collection Similar to Candidate variety Grouping of varieties To facilitate the assessment of distinctness, varieties to be divided into groups based on grouping characteristics. It is recommended that the testing organizations / centers use the following characteristics for grouping varieties (Maize): a) Tassel : Time of anthesis (cha # 4) b) Ear : Time of silk emergence (cha # 11) c) Ear : Anthocyanin coloration of silks (cha # 12) d) Plant : Length (cha # 15.1/15.2) e) Ear : Type of grain (cha # 22) UPOV, 2009 18

21. Characteristics and Symbols To assess distinctness, uniformity and stability, the characteristics and their states should be used. Notes (scale 1 to 9) should be use for the purpose of electronic data processing. Legend : (*) Characteristics that should be observed during every growing season on all varieties , included in the variety descriptions, except when the state of expression of a preceding characteristics show this inappropriate. (+) Characteristics is illustrated by explanation or drawing. (S) Possible segregation in three way cross and double cross hybrids. PPV & FRA, 2007 19

22. Table - 6: Range of characters as per PPV & FR Act Crop No. of characters Characters with * mark Wheat 33 20 Rice 30 5 Sunflower 40 13 Peas 58 27 Maize 31 (41- as per UPOV) 8 (16) Onion 27 12 * Mandatory characters to record. 20

23. Table - 7: Decimal code for the growth stage Stage code General Description 00 Dry seed 12 2 leaves unfolded 14 4 leaves unfolded 51 Inflorescence just visible 61 Beginning of anthesis 65 Anthesis halfway 71 Caryopsis watery ripe 75 Medium milk 85 Soft dough 92 Caryopsis hard ( can no longer be dented by thumbed nail) 93 Caryopsis loosening day time. New Delhi (Anonymous 2007) The optimum stage of plant growth assessment of each characteristics is indicated by decimal code. 21

24. Technical Questionnaire A Performa containing technical questionnaire about the candidate variety is to be submitted by the applicant seeking plant variety protection. The applicant is asked: Name of species Applicant (name and address) Proposed denomination or breeder’s reference Information on origin, maintenance and reproduction Type of material State of expression for each of the grouping characteristics Most similar varieties and the difference of candidate variety from these varieties Any additional information which can help to distinguish the variety Resistance to pests and disease Any special condition required for examination of the variety Any other useful information Cont…. 22

25. Cont…… IX. Technical Questionnaire Reference Number (Not to be filled in by the applicant.) To be completed with every application made on candidate variety in accordance with specific Indian dust test guidelines for registration under PPVFR&FR Act, 2001. 1. Species Zea mays L. Maize 2. Applicant (Name and address): ( In concurrence with column 3 of form no 1.) 3. Proposed denomination of the (candidate variety as given in form no 1) 4. Information on origin, maintenance and reproduction of the variety. 4.1 Type of variety Inbreed line . [ ] Single cross hybrid. [ ] Three way cross hybrid [ ] Double cross hybrid [ ] Open pollinated variety [ ] 4.2 Variety details Field corn [ ] Pop corn [ ] Baby corn [ ] Waxy corn [ ] Quality Protein Maize [ ] High oil corn [ ] Fodder corn [ ] 4.3 Pedigree of proposed variety/line 4.4 Other information 5. Characteristics of the candidate variety to be given (The number in note column refers to the different states of given characteristics described in column 3. Tick out within the brackets, which based corresponds to the character expression of the candidate variety): 23

26. 6. Similar varieties and difference from these varieties: Characteristics in which similar variety is different* Denomination of similar variety State of expression of candidate variety *: in the case of identical states of expression of both varieties, indicate the difference in grade of expression. 7. Additional information , which may help to distinguish the variety Resistance to pests diseases (indicate the race etc.) Produce quality Special conditions for the examination of the variety Other Information 8. Authorization for release State of expression of similar variety UPOV 2009 Sr. No. Characteristics States Note (1 to 9) Example variety/ line Stage Assessment 7. Tassel: anthocyanin coloration of anthers (in middle third of main axis on fresh anther ) Absent 1 CM 400 65 Anthesis halfway VG ( visually in a group) Present 9 CM 145 HKI 161 24

27. Sr. No. Characteristics States Note (1 to 9) Example variety / line Stage Assessment 1. (+) Leaf: angle between leaf blade and stem (on leaf just above upper ear) Narrow (<45 o ) 1 HHM 2 61 Beginning of anthesis VG (visually in a group) Wide (>45 o ) 9 Vivek 5 Table -8: Characteristics for Leaf: Angle between leaf blade and stem (on leaf just above upper ear) Narrow 25 Wide 1 9

28. Sr. No. Characteristics States Note (1 to 9) Example variety/ line Stage Assessment 2. (+) Leaf: Attitude of leaf blade (on leaf just above upper ear) Straight 1 HHM 2 61 Beginning of anthesis VG (visually in a group) Droopy 9 HKI 323 Table - 9: Characteristics for Leaf: Attitude of leaf blade (on leaf just above upper ear) Straight Droopy 26 1 9

29. Sr. No. Characteristics States Note (1 to 9) Example variety/ line Stage Assessment 3. Stem: Anthocyanins colouration of brace roots Absent 1 HKI 163 65-75 Anthesis halfway Caryopsis watery ripe, Medium milk VS (visually in a single) Present 9 HKI 327T Table -10: Characteristics for Stem : Anthocyanins colouration of brace roots 27 1 9

30. Sr. No. Characteristics States Note (1 to 7) Example variety/ line Stage Assessment 4. (*) Tassel: Time of anthesis (on middle third of main axis, 50 % of plant) Very early (<45 D) 1 HKI 335 65 Anthesis halfway VG (visually in a group) Early (45-50 D) 3 HKI 1025 Medium (50-55 D) 5 HKI 323 Late (>55 D) 7 HKI 1126 Table -11: Characteristics for Tassel: Time of anthesis (on middle third of main axis, 50 % of plant) 28

31. Sr. No. Characteristics States Note (1 to 9) Example variety/line Stage Assessment 5. (+) Tassel : Anthocyanin colouration at base of glume Absent 1 HKI 1344 65 Anthesis halfway VS (visually in a single) Present 9 HKI 161 Table -12: Characteristics for Tassel: Anthocyanins colouration at base of glume 29 1 9

32. Sr. No Characteristics States Note (1 to 9) Example variety/ line Stage Assessment 6. Tassel: Anthocyanin colouration of glumes excluding base (in middle third of main axis ) Absent 1 HKI 209 65 Anthesis halfway VS (visually in a single) Present 9 CM 300 HKI 161 Table -13: Characteristics for Tassel: Anthocyanin colouration of glumes excluding base (in middle third of main axis ) 9 30 1 9

33. Sr. No. Characteristics States Note (1 to 9) Example variety/ line Stage Assessment 7. Tassel: Anthocyanin colouration of anthers (in middle third of main axis on fresh anther ) Absent 1 HKI 209 65 Anthesis halfway VG (visually in a group) Present 9 CM 145 HKI 161 Table -14: Characteristics for Tassel : Anthocyanin colouration of anthers (in middle third of main axis on fresh anther) 9 1 31 1 9

34. Sr. No. Characteristics States Note (1 to 9) Example variety/ line Stage Assessment 8. Tassel: Density of spikelet (in middle third of main axis ) Sparse 3 HKI 1126 65 Anthesis halfway VG (visually in a group) Dense 7 HKI 288-2 Table -15: Characteristics for Tassel: Density of spikelet (in middle third of main axis) 9 1 32 3 7

35. Sr. No. Characteristics States Note (1 to 9) Example variety/ line Stage Assessment 9. (*) (+) Tassel: Angle between main axis and lateral branches (in lower third of tassel) Normal (<45 o ) 1 CM 145 65 Anthesis halfway VG (visually in a group) Wide (>45 o ) 9 CM 140 Table -16: Characteristics for Tassel : Angle between main axis and lateral branches (in lower third of tassel) 33 1 9

36. Sr. No. Characteristics States Note (1 to 9) Example variety/ line Stage Assessment 10. (*) (+) Tassel: Attitude of lateral branches (in lower third of tassel ) Straight 1 HKI 193-1 65 Anthesis halfway VG (visually in a group) Curved 5 HKI 323,46 Strongly curved 9 HKI 163 Table -17: Characteristics for Tassel : Attitude of lateral branches (in lower third of tassel) 34 1 9 5

37. Sr. No. Characteristics States Note (1 to 7) Example variety/ line Stage Assessment 11. Ear: Time of silk emergence (50 % plants) Very early (<48 D) 1 HKI 335 65 Anthesis halfway VG (visually in a group) Early (48-53 D) 3 HKI 1025 Medium (53-58 D ) 5 HKI 323 Late (>58) 7 HKI 1126 Table - 18: Characteristics for Ear: Time of silk emergence (50 % plants) 35

38. Sr. No. Characteristics States Note (1 to 9) Example variety/ line Stage Assessment 12. (*) Ear: Anthocyanin coloration of silks (on day of emergence) Absent 1 HKI 1025 65 Anthesis halfway VG (visually in a group) Present 9 HKI 323 Table - 19: Characteristics for Ear: Anthocyanin coloration of silks (on day of emergence) 36 1 9

39. Sr. No. Characteristics States Note (1 to 9) Example variety/ line Stage Assessment 13. Leaf: Anthocyanin coloration of leaf sheath (below ear) Absent 1 HKI 163 71 Caryopsis watery ripe VS (visually in a single) Present 9 CM 300 Table - 20: Characteristics for Leaf: Anthocyanin coloration of leaf sheath (below ear ) 37 1 9

40. Sr. No. Characteristics States Note (1 to 7) Example variety/ line Stage Assessment 14. Tassel: Length of main axis above lowest side branch Short (< 20 cm) 3 HKI 1128 71 Caryopsis watery ripe MS (measurements in single) Medium (20-30 cm) 5 HKI 327 T Long (>30 cm ) 7 V 25 Table - 21: Characteristics for Tassel: Length of main axis above lowest side branch 38

41. Sr. No. Characteristics States Note (1 to 9) Example Variety / line Stage Assessment 15.1 (*) Inbred line only : Plant Length (up to flag leaf ) Short (<120cm) 3 HKI 1348-6-2 75 Medium milk MS (measurements in single) Medium (120-150 cm ) 5 HKI 323 Long (>150 cm) 7 HKI 1128 15.2 (*) Hybrids and open pollinated varieties only : Plant length (up to flag leaf) Short (<150cm) 3 Vivek 75 Medium milk MS (measurements in single) Medium (150-180 cm ) 5 Prakash Long (180-210 cm) 7 HQPM 1 Very long (>210 cm) 9 African Tall Table - 22: Characteristics for Plant Length (up to flag leaf) 39

42. Sr. No. Characteristics States Note (3 to 7 ) Example variety/ line Stage Assessment 16. Plant: Ear placement Low 3 HKI 1011 75 Medium milk MS (measurements in single) Medium 5 HM 4 High 7 HQPM 1 Table - 23: Characteristics for Plant: Ear placement 40

43. Sr. No. Characteristics States Note (3 to7) Example variety/ line Stage Assessment 17. Leaf : Width of leaf blade (leaf of upper ear) Narrow(<8cm) 3 HKI 323 75 Medium milk MS (measurements in single) Medium(8-9cm) 5 HKI 295 Broad (>9 cm) 7 HKI 1126 Table - 24: Characteristics for Leaf : Width of leaf blade (leaf of upper ear) 41

44. Sr. No. Characteristics States Note (3 to 7) Example variety/ line Stage Assessment 18. (*) Ear: Length without husk Short (<10cm) 3 HKI 536 92 Caryopsis hard MS (measurements in single) Medium (10-15 cm) 5 HKI 163 Long(>15 cm) 7 HQPM 1 19. Ear: Diameter without husk (in middle) Small (<4 cm) 3 HKI 323 92 Caryopsis hard MS (measurements in single) Medium (4-5 cm) 5 HKI 327 Large (>5 cm) 7 THQPM 1 Table - 25: Characteristics for: Ear length without husk and Ear diameter without husk (in middle) 42

45. Sr. No. Characteristics States Note (1 to 3) Example Variety / line Stage Assessment 20. (+) Ear shape Conical 1 HKI 1344 92 Caryopsis hard VG (visually in a group ) Conical-Cylindrical 2 HKI 295 Cylindrical 3 HKI 1105 Table - 26: Characteristics for Ear Shape 43 1 3 2

46. Sr. No. Characteristics States Note (3 to 7) Example variety/ line Stage Assessment 21. Ear : Number of row of grains Few (≤ 8) 3 - 92 Caryopsis hard MS (measurements in single) Medium (10-12) 5 HKI 163 Large (≥ 14) 7 HM 5 Table - 27: Characteristics for Ear : Number of row of grains 44 L-16 row, R-14 row

47. Sr. No. Characteristics States Note (1 to 3) Example variety/ line Stage Assessment 22. (*) Ear : type of grain (in middle third of ear) Flint 1 HKI 1105 92 Caryopsis hard VG (visually in a group ) Semi flint/ Semi dent 2 HKI 1344 Dent 3 HM 5 Table - 28: Characteristics for Ear : Type of grain (in middle third of ear) 45 1 3 2

48. Table - 29: Characteristics for Ear : Colour of top of grain Sr. No. Characteristics States Note (1 to 7) Example variety/ line Stage Assessment 23. (*) Ear: Colour of top of grain White 1 CM 300 92 Caryopsis hard VG (visually in a group ) White with cap 2 HKI 1344 Yellow 3 HKI 1050 Yellow with cap 4 HKI 209 Orange 5 HKI 323 Red 6 Variegated 7 2 3 5 6 46 4 1

49. Table - 30: Characteristics for Ear : Anthocyanin colouration of glumes of cob Sr. No. Characteristics States Note (1 to 3) Example variety/ line Stage Assessment 24. (*) Ear: Anthocyanin colouration of glumes of cob White 1 HKI 163 93 Caryopsis loosening day time VG (visually in a group ) Light purple 2 SUWAN Dark purple 3 HKI 161 1 2 3 47

50. Table - 31: Characteristics for Kernel : Row arrangement (middle of corn) Sr. No. Characteristics States Note (1 to 3) Example variety/ line Stage Assessment 25. (*) Kernel : Row arrangement (middle of corn) Straight 1 HM 2 93 Caryopsis loosening daytime VG ( visually in a group ) Spiral 2 HM 5 Irregular 3 HKI 1344 48 1 2 3

51. Table - 32: Characteristics for Ear : Type of grain Sr. No. Characteristics States Note (1 to 5) Example variety/ line Stage Assessment 26-29. Ear : Type of grain Field corn 1 CM128 92 Caryopsis hard VG (visually in a group ) Pop corn 2 VL. Popcorn Waxy / opaque 3 Vivek Hy.4 Opaque tinge 4 Vivek Hy.5 Sweet corn 5 Madhuri 1 5 49 1 2 3 4

52. Sr. No. Characteristics States Note (1 to 5) Example variety/ line Stage Assessment 30. (+) Kernel shape Shrunken 1 Madhuri 93 Caryopsis loosening day time VG (visually in a group ) Round 2 HKI 1342 Indented 3 HM 5 Toothed 4 HKI 1348-6-2 Pointed 5 - Table - 33: Characteristics for Kernel shape PPV & FRA, 2007 5 50 3 4 2 1

53. Table - 34 : Characteristics for Kernel size (1000 kernel weight) Sr. No. Characteristics States Note (1 to 7) Example variety/ line Stage Assessment 31. Kernel size (1000 kernel weight ) Very small (<100g) 1 Madhuri 93 Caryopsis loosening day time MG (measurements in a group ) Small (100-200 g) 3 HKI 1025 Medium (200-300g) 5 Prabhat Large (>300g) 7 V.HY.-9 51

54. Conclusions India has the largest reservoir of the genetic material and accounts for the largest skilled population in the world, it gives several opportunities to the Indian agricultural research and development. To validate the claims distinctness, uniformity and stability (DUS) tests will have to be conducted as per the National Test Guidelines. PPV & FR Act protects breeders varieties from unauthorized infringement, ensure farmers rights on seeds and encourage growth of seed industries by making seeds of protected varieties available to the farmers. DUS Testing is one of the key feature of plant variety protection. For easy identification and preservation DUS test is very important . 52

55. Future thrust DUS test should be modified according to need, cropping pattern and environmental conditions. More emphasis should be given on DUS testing by scientists / technical qualified persons . The institutes/ research organization engaged in plant breeding should stream line their maintenance breeding programme so as to make available pure seed, which shall be used for DUS tests and kept as reference sample for at least 7 years. A well conceived programme of training of staff for conduct of DUS tests, data compilation, analysis and its interpretation should be organized. 53

56. 54 Thank You