Table 41. Results of breeding efforts, 2015.
Nursery Number of entries evaluated
Number of entries selected
Remarks Parentals 160
varieties/lines
160 varieties/lines Most of these entries have the desired quality characteristics, yield enhancing traits and resistance specifically for blast, BLB and tungro New
crosses
- 99 Aromatic (A), 53
Glutinous(G), 44 Pigmented(P) and 66 Zn/Fe
Seed setting of the crosses made was affected by inclement weather condition in WS
F1 (crosses)
156A, 63G, 62P and 121 Zn/Fe
91 A, 29 G, 17 P and 89 Zn/|Fe for HPS 7A, 15G, 2 P for generation advance
Other crosses were discarded owing to poor plant type and susceptibility to BLB and blast
11A, 17P and 4Zn/Fe were stored in the cold room to be planted in 2016DS
MBN (Hybrid populations for generation advance)
77A, 20G, 24P, 80Zn/Fe
33A, 6G, 10P and 26Zn/Fe for plant selection
16A, 5G, 4P, 25Zn/Fe retained for generation advance
In the DS,13A, 6P and 5Zn/Fe were discarded owing to poor plant type, severe lodging and susceptibility to BLB, sheath blight and blast.
Selection during the WS was also limited due to severe lodging and germination of the plants brought by two typhoons that visited the area when the plants were at soft dough (Kabayan) and hard dough/maturity (Lando).
HPS (Hybrid populations intended for single plant selection)
58A, 29G, 30P, 15Zn/Fe
Field selection= 12982 individual plants Laboratory = 3661 plants
1673A, 1480G, 1140P, 296 Zn/Fe
Selection during the WS was also limited due to severe lodging and germination of the plants brought by two typhoons that visited the area.
Pedigree Nursery (PNG)
3110 pedigree lines (1603A, 635G, 295P 577 Zn/Fe)
Final selection= 795 A, 319 G, 153 P, 260 Zn/Fe.
Of these, 55A, 19G, 3P and 21 Zn/Fe uniform lines were elevated to AON for 2015 WS planting
Selection during the WS was also limited due to severe lodging and germination of the plants brought by two typhoons that visited the area.
Figure 27. Kernel quality of the selected plants per specialty types from the HPS and PN, 2015DS.
Figure 28. Selected plants in the hybrid populations (HP) and pedigree nursery (PN).
Promising lines in the performance trials
• In the Advanced Observation Nursery (AON), out of the 247 entries evaluated, 36 ( A-21, Zn/Fe-5, P-10) were retained for further evaluation and 67 (A-24, Zn/Fe-19, G-10, P-14) were elevated to Preliminary Yield Trial (PYT). There were 31 non- aromatic entries but with high yields ranging from 6292kg/
ha to 9589kg/ha and with good to excellent kernel qualities were transferred to AON-IL (18) and PYT-IL (13). Figure 29 shows the yield performance of the ten top yielding entries per specialty types elevated to (PYT). Yield range involving these entries were 7496kg/ha-8674kg/ha for A, 7219kg/ha-
9738kg/ha for Zn/Fe, 6962kg/ha-9635kg/ha for P and 5638kg/
ha-9294kg/ha for G. These entries showed better yield performance than the corresponding group check variety.
• Promising entries were also identified among 130 PYT entries.
Of these, 59 were retained for further evaluation. There were 2 A, 3 Zn/Fe, 1 P elevated to the multi-location yield trial (MYT) 2015WS evaluation in CES awaiting slots in the MET. Seed samples of these test entries were submitted to the Rice Chemistry and Food Science Division for grain quality evaluation. Figure 30 shows the other promising test entries with comparable to or better yield performance than the corresponding specialty type check variety.
Figure 29. Yield performance and maturity of the top yielding entries per specialty types in the AON, 2015DS.
Figure 30. Yield performance and maturity of the top yielding entries per specialty types in the PYT, 2015DS.
Promising lines in the multi-location trials
• In the most advance stage of Multi-Environment Yield Trial (MET, Stage 2 module 2), there were 22 entries evaluated during the season. PR38168-2B-3-1-3-1-1-1-2, an aromatic line, ranked 7th across 7 test locations with an average yield of 5.43t/ha. This entry ranked 2nd in PhilRice San Mateo and 3rd in PhilRice Agusan with yields of 7.64t/ha and 5.79t/ha., respectively. Its AMMI Stability Values (ASV) across sites was 0.57 and ranked 12th majority of the test entries yielded comparably with the while its Yield Stability Index (YSI) was 19 and ranked 5th among 22 test entries.
PR38963-(Fe)-B-5-4-2-1-1, another entry in the same group, ranked 10th with an average yield of 5.40t/ha. It registered the highest yield (7.71t/ha) in PhilRice San Mateo and 2nd in PhilRice CES with 9.06t/ha yield.
• PR38012-3B-1-3, a promising aromatic line which completed the required trials in MET was elevated to NCT for Special Purpose (NCT-SP), 2015W.
• In the NCT-SP, out of the 8 A, 8 G and 8 Zn/Fe test entries, there were 4 A, 7 G and 2 Zn/Fe PhilRice developed lines under evaluation. Under Maligaya condition, three A entries showed numerically higher yield performance than the aromatic check, NSIC Rc128 (9.9t/ha). These were PR38949-B-29-2 (10.7/ha), PR36905-B-1-11-2-1-1-1(10.7t/
ha) and PR36914-B-9-4-1-6-4-1(10.3t/ha). These entries ranked 2nd, 3rd and 4th, respectively. Among the glutinous entries, PR34859-B-4-1-1-2-1 was the top yielder (10.8t/
ha) with 9.0% yield advantage over NSIC Rc13 (9.9t/ha), the highest yielding check. PR41035-B-B-17-2-3-1 (9.4t/
ha), another G entry, outyielded NSIC Rc15 (8.6t/ha) by 9.3%. Among the micronutrient-dense test entries, PR38963 (Fe)-B-7-1 (10.2t/ha) ranked 2nd with 20% yield advantage over PSB Rc82 (8.5t/ha), the yield check and 50% over MS13 (6.8t/ha), the micronutrient check variety.
Evaluation and utilization of fragrance markers for high-yielding aromatic breeding materials
TE Mananghaya, MV Embate, KC Geneston, HT Ticman , EC Arocena and LM Perez
Fragrance (aroma) is considered one of the most important grain quality traits in rice because aromatic rice cultivars command premium prices in the market today. However, the trait is recessive and thus breeding of aromatic rices needs extensive labor and has to involve large numbers of breeding materials to develop and identify a progeny with the target aromatic trait. Thus, molecular markers associated with fragrance trait will assist rice breeders to develop a high yielding variety with excellent grain quality in simple and inexpensive method. This study aims to evaluate a marker system to be used in marker assisted selection for fragrance trait in rice, and thus in aromatic breeding program.
Highlights:
• Assembled and evaluated new functional markers, the FMbadh2-E2A, FMbadh2-E2B and ous and 143 plants were negative to fragrance gene. The F2 plants detected positive to fragrance gene and found aromatic trait using KOH method were selected for forward breeding.
• Polymorphism survey using multiplex fragrance markers were conducted in 102 germplasm materials composed of 96 Philippine landraces and 6 improved varieties. Black Rice labelled in North Cotabato for its export quality was detected negative to fragrant allele, as well as Malagkit Sungsong and Improved Malagkit Sungsong (IMS-2). NSIC Rc128, NSIC Rc342 and NSIC Rc344, released rice varieties claimed aromatic also showed homozygous non fragrant allele using the diagnostic markers. While, NSIC Rc148, NSIC Rc218, Asucena and Binarit showed diagnostic allele for homozygous fragrant allele (Figure 31). Out of 96 traditional varieties evaluated, 74 F2 plants were identified positive to fragrant allele. These potential parents can be used as pollen donor for aromatic rice breeding in PhilRice.
• Segregating population of PR40476-3B-1-1/HHZ12-DT10- SAL1-DT1, PR37994-B-20-2-2-1/PR34627-B-44-2-1-2-1-1, HHZ3-SAL13-Y1-SAL1/PR37343-B-6-3-2-2-2 were established for evaluation of fragrance trait. For each cross combination, 200 F2 plants were individually tagged and leaf samples were collected prior to DNA genotyping using fragrance markers. A total of 600 F2 plants were evaluated and 284 plants showed fragrant allele, 112 detected heterozygous and 143 plants
were negative to fragrance gene. The F2 plants detected positive to fragrance gene and found aromatic trait using KOH method were selected for forward breeding.
Figure 31. Result of DNA analysis of selected parents and traditional rice cultivars using fragrance markers (Bradbury 2005).
Table 42. Summary of DNA genotyping of F2 segregating plants for detection of fragrant trait using multiplex PCR fragrance markers.
Field Code
2015 DS Parentage Pedigree Name Generation Fragrant Non-Fragrant Heterozygous
HPS-SP-1-1 PR40476-3B-1-1/HHZ12-DT10-SAL1-DT1 PR46405-B-1 F2:3 157 21 24
HPS-SP-5-1 PR37994-B-20-2-2-1/PR34627-B-44-2-1-2-1-1 PR46409-B-5-1 F2:3 64 71 46
HPS-SP-6-1 HHZ3-SAL13-Y1-SAL1/PR37343-B-6-3-2-2-2 PR46416-B-6-1 F2:3 63 51 42
Fragrance Genotyping
Production of golden rice introgression lines in the background of selected popular varieties with resistance to tungro and bacterial leaf blight
CC Adeva, RT Miranda, CFS Te, NR Sevilla and AA Alfonso
Golden Rice (GR), which contains elevated levels of the pro-vitamin A beta-carotene in the grains, was developed about a decade ago through biochemical engineering of the carotenoid biosynthetic pathway. It is eyed as an additional food-based strategy to combat the pervasive and persistent problem of vitamin A deficiency (VAD) especially in most rice-eating countries. VAD can damage the immune system and decrease the body’s ability to resist or fight infections; thereby, increasing the risk of mortality from common diseases, especially among women and young children. It may also result in impaired vision, including night blindness and permanent blindness. In the Philippines, about four out of ten children aged six months to five years and three out of every ten school-aged children have VAD. In addition, one out of every five pregnant and one out of every five lactating mothers have VAD. VAD continues to adversely affect many people despite nutrition interventions such as vitamin A fortification and supplementation programs that seek to provide adequate vitamin A to those at risk. Further research has elevated the amount of vitamin A to a meaningful level (up to 36 µg/g) and preliminary results indicate high conversion rate (4:1) of beta-
carotene from GR into retinol. The Golden Rice donor is a US commercial variety that grows poorly under Philippine condition. This projects aims to develop Golden Rice adapted under Philippine condition.
Highlights:
• Phenotypic and genotypic evaluations were done to characterize the identified 2-in-1 line donor, PR37171-1-1- 1-2-2-1-1 (also designated as LINE 27) and NSIC Rc120, also known as Matatag 6 (tungro resistant donor).
• LINE 27 and NSIC Rc120 were both phenotypically screened for Rice Tungro Virus (RTV) and Bacterial Leaf Blight (BLB) resistance. Table 43 shows that Line 27 has intermediate reaction while NSIC Rc120 has resistance reaction against RTV. For BLB screening, LINE 27 was confirmed to have high resistance reaction against Race 3 (Maligaya Race) and Race 6, the most virulent Xanthomonas oryzae pv. oryzae (Xoo) strain;
while NSIC Rc120 shows resistance reaction only to Race 3.
• Line 27 and NSIC Rc120 along with ARC11554 (positive check) and Taichung Native 1 (TN1) (negative check) were genotypically evaluated using Simple Sequence Repeat (SSR) markers RM5495 and RM8213 linked to rice tungro spherical virus 1 (tsv1) loci for tungro resistance and Green leafhopper resistance-14 (Glh14) loci for green leafhopper (GLH) resistance. Figure 32 shows that LINE 27 has similar expected band size to TN1 which is susceptible to tungro spherical virus but resistant to green leafhopper which is similar to the expected band size of ARC11554. On the other hand, NSIC Rc120 has similar expected band size to ARC11554 possessing both resistance to tungro spherical virus and green leafhopper.
• Two F2 populations of reciprocal crosses between LINE 27 and NSIC Rc120 were generated and advanced. In addition, four new F1s were generated with one backcross and three double crosses between F1 and LINE 27.
Table 43. Phenotypic reactions of different lines to Races 3 and 6 of Xoo at 14-days after inoculation and to tungro resistance at three to four weeks after inoculation.
ENTRY CODE
Race 3 Race 6
RTV Rating Lesion length
(cm) BLB Rating Lesion Length
(cm) BLB Rating
Line 27 0.2 R 0.1 R I
NSIC Rc120 2.57 R 31.9 S R
ARC11554 - - 6.2 I R
UTRIMERAH - - 16.7 S R
IR64 - - - - MS
TN1 33.8 S 30.9 S S
IR24 10.4 MS 29.03 S -
IRBB4 11.8 MS 11.7 MS -
IRBB5 12.3 MS 11.9 MS -
IRBB7 18.4 S 19.6 S -
IRBB21 9.49 I 11.9 MS -
IRBB61 3.37 R 8.2 I -
Legend: RTV Rating: R – resistant; MS – moderately resistant; I – Intermediate; S –Susceptible Lesion length/BLB Rating: 0-5 cm (R-Resistant); >5-10 cm (I-Intermediate); >10-15 cm (MS Moderately susceptible); >15 cm (S-Susceptible)
Figure 32. PCR amplification of LINE 27 (PR37171-1-1-1-2-2-1-1) and NSIC Rc120 (Matatag 6) together with resistant (ARC11554) and susceptible (TN1) checks for rice tungro virus using functional SSR markers: RM5495 (tsv1 for tungro resistance) and RM8213 (Glh14 for GLH resistance). L = DNA marker
(100 bp).