RESULT AND DISCUSSION 1 Growth and genetic parameters

Bogor, 21-22 October 2015 765 ………λλλλλλλλλλλλλλ. 5 where, b j is the coefficient of weight for the j-th trait, and x ij is the adjusted least square estimate of the i-th family on the j-th trait. The coefficient of weight b was calculated as a form vector by the following matrix formula: λλλλλλλλλλλλλλλ..λλ..λλλλλλλλλ6 where, P, G and a are phenotypic variance covariance matrix, that of family and vector of relative economic weight for the traits, respectively. The adjusted least square estimate was calculated by the following formula: [ ⁄ ] ⁄ ........................................................................................7 where, a ij , σ 2 fj , σ 2 ej and n i. are the lest square estimate of the i-th family on the j-th trait, the family variance component of the j-th trait, that of error variance and number of plots of the i-th family, respectively. For the convenience of analysis, expected genetic gain brought by family selection based on the index value is calculated by assuming one unit of selection intensity with the formula as follows: [ ] λλλλλλλλλλ.λλλ..λλλλλλ..λλλ8 Comparisons for variances, genetic parameters and genetic gain between the two groups of family: type-1+2 and type-1+2+3 were made to observe the impact of “recycled genetic resources” in the third-generation progeny trial of A. mangium. 3. RESULT AND DISCUSSION 3.1 Growth and genetic parameters Along the first three year ages, growth and form traits varied among the three Types of family Table 1. Plus trees families from Type-1 which were improved through two cycles of breeding generally performed better than the families from Type-2 and Type-3 in all traits with superiority ranged from 0.8 - 9. Subsequently the Type-2 were generally better than Type- 3, except for stem straightness. This result indicated that the series of selection practiced in the first and second-generation breeding cycles were done properly to achieve higher positive genetic gain accumulatively from generation to generation. The superiorities resulted in this study were in agreement with the report of realized genetic gain observed in second-generation seedling seed orchard Nirsatmanto et al., 2004; Nirsatmanto et. al., 2013. It should be noted here that the superiority is a response to family and plus trees selection in the orchard and the gain seemed to become greater in operational if compared to the unimproved seed. This is because the sub-line B used in this study consisted of family originated from Oriomo region which was recognized as the most productive of PNG provenances Harwood Williams, 1991, Kari et al., 1996. Bogor, 21-22 October 2015 766 Combining 57 families from all Types Type-1+Type-2+Type-3 resulted similar family variation as those combining from only 44 families Type-1+Type-2, except for height at two years of age. This similar family variation seemed due to an identical by descent in family between Type-1 and Type-3. This is because selected family in Type-3 added into third- generation is same as those in Type-1. As a result, the magnitude of family variation was maintained similarly between both groups of type family 57 families and 44 families. On the other hand, however, the magnitude of variation from the 44 families was accompanied by increasing of their error variance. This is indicated that the reduction number of families is subject to greater experimental error, even though the increasing of error variance was not reflected in the estimates of heritability. The value of heritability was more reflected by the magnitude of family variance. Table 1: Mean of height, dbh, stem straightness and genetic parameters from three years of age measurements Family and genetic parameters height m dbh cm stem straightness age-1 age-2 age-3 age-2 age-3 age-2 age-3 Type-1 2.50±0.03 7.19±0.06 11.88±0.05 6.7±0.09 11.4±0.12 3.6±0.03 3.6±0.04 Type-2 2.48±0.04 7.07±0.08 11.66±0.06 6.3±0.12 10.8±0.15 3.3±0.05 3.4±0.05 Type-3 2.45±0.04 6.95±0.09 11.57±0.07 6.0±0.12 10.6±0.18 3.5±0.05 3.6±0.05 Grand mean 2.48±0.02 7.11±0.04 11.75±0.03 6.4±0.06 11.1±0.08 3.5±0.02 3.5±0.02 Family variance component 2 f 0.0126 0.0155 0.0791 0.0633 0.0900 0.0904 0.3126 0.3173 0.5203 0.5213 0.0134 0.0186 0.0345 0.0411 Error variance 2 e 0.2396 0.2618 0.9999 1.0274 1.1351 1.1456 2.8162 3.0069 6.5981 6.6526 0.7018 0.6937 0.7755 0.8086 Individual heritability h 2 i 0.13 0.15 0.18 0.14 0.27 0.27 0.32 0.30 0.27 0.27 0.07 0.09 0.15 0.17 Family heritability h 2 f 0.26 0.29 0.31 0.27 0.54 0.54 0.52 0.51 0.54 0.54 0.21 0.25 0.37 0.41 Note: number in parentheses is the parameter that was calculated based on combination of 44 families from type- 1 and type-2 only type-3 excluded Table 2: Observed significance associated with the analysis of variance for three years measurements of height, dbh and stem straightness Source of variance p-value height year dbh year Straightness year age-1 age-2 age-3 age-2 age-3 age-2 age-3 Replications .0001 .0001 .0001 .0001 .0001 .0001 .0001 .0001 .0001 .0001 .0001 .0001 .0001 .0001 Type of family 0.4317 0.4641 0.0043 0.0361 0.0001 0.0029 .0001 0.0005 0.0001 0.0029 .0001 .0001 0.0012 0.0035 Family within Type .0.0001 .0001 .0001 .0001 .0001 .0001 .0001 .0001 .0001 .0001 0.0939 0.0366 .0001 .0001 Family x replication .0001 .0001 .0001 .0001 0.0093 0.0233 .0001 .0001 0.0093 0.0233 0.0006 0.0001 0.0004 0.0060 Note: number in parentheses is the observed p-value based on combination of 44 families from type-1 and type-2 only type-3 excluded Bogor, 21-22 October 2015 767 Adding the Type-3 into third-generation breeding population tended to increase the family variation, but the magnitude of variation seemed to be reduced as tree getting older Table 2. Except for stem straightness at two years of age, variation of the family within the types showed a similar significance among the 57 families and 44 families. It confirmed a consistently significance of the family variations among the two groups along the three year measurements, except for stem straightness at two years of age. This result indicated that the amount of genetic improvement could be continuous attained through third-generation of tree improvement. In addition, the increased of error variance in 44 families group was not reflected into the weakness of significances. Correlations within growth traits height and dbh using the data from 57 families were higher than the correlation between growth traits and form trait Table 3. The correlations between the growth traits and form trait were generally weak, and for some cases were negative. On the other hand, genotypic correlation was consistently higher than phenotypic correlation for almost all traits, indicating that stronger relationships existed in genetic than that in phenotype. The high genetic correlation accompanied with high estimates of heritability would become essential factor for observing the family performances using selection index. This is because the index would be constructed by combining information from several traits. The value of genetic correlation and heritability obtained in this study seemed to be sufficient for constructing the selection index for A. mangium, especially for the traits at two years of age. On the other hand, the weak and negative correlation based on the data in three years of age brought the data from this age would not sufficiently available for selection index analysis. Table 3: Phenotypic correlation bellow diagonal and genotypic correlation above diagonal among the measured traits from 55 families Traits height-1 height-2 height-3 dbh-2 dbh-3 straightness-2 straightness-3 height-1 - 1.0285 0.7668 0.6526 0.7612 0.5710 0.6798 height-2 0.7940 - 0.9089 0.9926 0.9013 0.4658 0.3184 height-3 0.6769 0.7748 - 0.9452 1.0000 0.3310 -0.1316 dbh-2 0.7049 0.8630 0.9023 - 0.9450 0.3347 -0.0912 dbh-3 0.6746 0.7717 0.9999 0.9020 - 0.3187 -01365 straightness-2 0.2007 0.2492 0.2066 0.1853 0.2032 - 0.6874 straightness-3 0.1144 0.0971 -0.1077 -0.1407 -0.1099 0.410 - 3.2 Selection index In current study, selection index was constructed based upon the data from two years of age instead of the data from three years of age, involving tree height, dbh and stem straightness. This is because the weak of correlation among the traits at three years of age, especially for stem straightness. Due to the difficulty and limitation of the available data, for convenience the relative economic weight value was calculated as an inverse of the phenotypic standard deviation of family mean in each trait. Among the traits used for index calculation, growth traits height and dbh received higher coefficient weight than form trait stem straightness in the both groups of family type Table 4. The lower weight for stem straightness might reflect the low of family variance as indicated by the weak and non-significant differences among the tested families Table 2. By assuming the same number of selected family 22 families, that is intensity of selection at 1.00 for Group 1 and at 0.789 for Group 2, the selection index provided a positive value of selection differential for all traits. The highest selection differential was found at dbh, which was then followed by tree height and stem straightness. Except for stem straightness, selection Bogor, 21-22 October 2015 768 differential for family in Group 1 was higher than that in Group 2. Although the mean value of each trait was lower, absolute gain in Group 1 was higher than that in Group 2 and it provided increases of relative genetic gain at around 15 - 25 for tree height and dbh. It indicated that adding some amount of number family from the original founder families in first generation grandparents increased genetic gain in third-generation breeding cycles, especially for the traits which having high heritability. Table 4: Summary of coefficient weight and expected genetic gain based on selection index at the two years of age from matrix analysis Group Family Trait b Selection Differential 1 Absolut e gain 1 Mean 1 Relative gain Type Number 1 Type-1 + 2 + 3 57 Height-2 0.863 0.533 0.209 7.031 3.0 Dbh-2 0.849 0.789 0.351 6.364 5.5 Straightness-2 0.409 0.064 0.030 3.473 0.9 2 Type-1 + 2 44 Height-2 0.696 0.423 0.171 7.068 2.4 Dbh-2 0.717 0.642 0.309 6.465 4.8 Straightness-2 0.567 0.086 0.031 3.461 0.9 Note: 1 In meter unit for height-2 and centimeter unit for dbh-2. Intensity selection was set up at the same number of selected family 22 families: I=1.00 for Group 1 and I= 0.798 for Group 2. Table 5: Family ranking of the three types of family determined using selection index basedn on the traits at two years of age Type-1 Type-2 Type-3 Family no. Index Rank Family no. Index Rank Family no. Index Rank 1 14.276 9 28 13.278 23 45 12.952 27 2 13.929 12 29 11.640 50 46 11.871 46 3 14.405 6 30 14.436 5 47 12.017 45 4 13.324 21 31 11.497 52 48 12.817 29 5 13.589 16 32 12.215 42 49 12.496 35 6 12.486 36 33 12.399 37 50 11.426 53 7 12.917 28 34 12.665 30 51 11.746 49 8 12.269 40 35 13.310 22 52 11.575 51 9 14.527 4 36 13.165 25 53 13.450 19 10 14.951 2 37 10.126 57 54 11.819 48 11 13.511 18 38 10.965 55 55 14.367 7 12 14.111 11 39 12.530 33 56 12.347 38 13 13.178 24 40 14.559 3 57 13.886 14 14 11.336 54 41 11.840 47 15 12.324 39 42 13.334 20 16 12.507 34 43 13.596 15 17 12.581 32 44 12.060 44 18 14.151 10 19 12.267 41 20 10.690 56 21 12.198 43 22 12.972 26 23 14.351 8 24 12.600 31 25 13.920 13 26 13.555 17 27 15.500 1 Bogor, 21-22 October 2015 769 The increased of genetic gain in Group 1 compared to Group 2 was mainly due to the increasing number of high quality plus trees family, which is some of them coming from the Type-3 Family. According to the family ranking generated using selection index value, out of 13 families from Type -3 that was added into the third-generation progeny trial around 23 families of them classified into the top twenty family ranking, followed by 30 into the middle twenty and 53 into the lowest ranking Table 5. These numbers of families are equivalent to the 15 of the total top twenty family ranking, 25 of the total middle twenty and 35 of the total lowest family ranking Figure 1. Figure 1: Proportion number from three types of families classified based on the family ranking Among the top twenty family rankings, the ranks value of family from Type 3 were 5 - 65 superior to the ranks value of family from Type-1 and Type-2. It means that among the top ranking, selected families from Type-3 is not always poorer compared to those from Type-1 and Type-2, even they showed more productive in some cases. This result revealed that the uses of within-family variation is potential for A. mangium breeding. This is one of the essential factors to apply the “recycled genetic resources” for advanced-generation breeding of A. mangium. 3.3 Pedigree control for advanced generation breeding Maintenance of pedigree control is critical in a breeding population. The pedigree control would become more complex as the advanced generation breeding cycles progressed. Moreover the increase of relatedness in advanced generation population coupled with the near-universal importance inbreeding depression in forest trees lead the management of inbreeding would become a key issue in the development and implementation of all advanced-generation breeding program White, 1996. Therefore generating genetic structure through controlling the pedigree should be taken into account in order to maintain the genetic diversity and to achieve the long term genetic gain. One of the rules for achieving a great genetic gain while maintaining genetic diversity in advanced-generation breeding population is the limitation maximum number of plus trees within each selected family. In this study, maximum two plus trees per family is a τrule of thumb” applied for controlling the pedigree in establishing advanced-generation breeding of A. mangium. Therefore identical by descent of family should be handled carefully in advanced 10 20 30 40 50 60 70 Type-1 Type-2 Type-3 N u m b e r o f Fa m il ie s Types of Family Total Top-20 Middle-20 Lowest-17 Bogor, 21-22 October 2015 770 generation breeding program. Concerning the need of pedigree control as described above, the “recycled genetic resources” as proposed in this study that is a re-uses of the original founder families of first generation grandparents into third-generation breeding population, could be adopted with some necessarily requirements as follows: a. Families from grandparent population founder families should be selected based on the result of family selection in second-generation breeding population, for this, pedigree of the selected families should be always confirmed back to the original parents or ancestors Figure 2. b. Selected family from grandparent population founder families could be inserted into third- generation breeding population only if the number of plus trees for respective family and their identical by descent family selected in the second-generation is less than two plus trees. As a result the total maximum number of plus tree per family is finally only two, including the plus trees family from grandparent population. Figure 2: Flow chart of pedigree control from certain family ex. No. family 100 into their successive generations up to third-generation Based on the requirements as described in preceding paragraph, the “recycled genetic resources” would be an alternative method to optimize the genetic resources for A. mangium advanced generation breeding program. This is because the genetic diversity of A. mangium was reported as low, especially diversity within population Butcher et al., 1998. Concerning the important of large genetic diversity for achieving long term genetic gain, infusion from new selection families in natural forest would be one of the solutions. However, recently the availability of such infusion families was much reduced and not always available either in quantity or in quality. The results of this study revealed that the τrecycled genetic re source” through re-uses of the original founder families of first generation grandparents into third-generation breeding population is applicable for maintaining genetic diversity and increasing genetic gain of A. mangium. However, pedigree control of the families should be taken into account to reduce inbreeding incidence in the population. Bogor, 21-22 October 2015 771

4. CONCLUSION