HERITABILITY AND HETEROSIS OF GRAIN YIELD ON DOWNY MILDEW RESISTANCE (DMR) AND QUALITY PROTEIN MAIZE (QPM) INBREDS

AND THEIR SINGLE CROSS HYBRIDS

(HERITABILITAS DAN HETEROSIS HASIL PADA GALUR-GALUR DMR DAN QPM SERTA HIBRIDA SILANG TUNGGALNYA)

Ruswandi, D.1), N. Wicaksana2), M. Rachmadi1), A. Ismail2), D. Arief2), and F. Kasim3) Kata kunci : heritabilitas, heterosis, Perenosclerospora maydis, jagung berkualitas protein tinggi. Key words : heritability, heterosis, Perenosclerospora maydis, quality protein maize.

Abstract
Improvement of high yield maize cultivars possessing high quality of seed as showed by high lysine and tryptophan content must be fulfilled with resistance against downy mildew pathogen in any maize breeding program. A set of experiment to study genetic variability and to estimate both heritability and heterosis of important characters of grain yield on DMR and QPM lines has been conducted. Four downy mildew resistance (DMR) lines, i.e. Nei 9008, P 345, Ki 3, and MR 10; three quality protein maize (QPM) lines, including CML 161, CML 163, and CML 172; and their single cross hybrids formed based on line tester mating design were used in the experiment. The following characters: height of the first ear, seeds number per row, seed weight per ear, and grain yield per plot showed broad genetic variability. Narrow sense heritability (hns) of grain yield and its components ranged from 0.01 to 0.71. Broad
1) Assistant Professor, Lab. of Plant Breeding, Department of Crop Sciences, Padjadjaran University, Bandung. Contact email: ruswandi@hotmail.com 2) Research Assistant, Lab. of Plant Breeding, Department of Crop Sciences, Padjadjaran University, Bandung. 3) Maize Breeder, Indonesian Cereals Research Institute (ICERI), Maros

sense heritability (hbs), on the other hand, ranged from 0.20 to 0.74. It was found that hybrid derived from crossing between parental inbred lines possessing broad genetic background would express high heterosis.

Sari
Pengembangan kultivar jagung berdaya hasil tinggi yang memiliki kualitas biji yang tinggi, yang ditandai oleh tingginya kandungan lisin dan triptofan harus diikuti dengan ketahanan terhadap pathogen bulai pada setiap program pemuliaan jagung. Satu set penelitian untuk mempelajari variabilitas genetik dan untuk menduga heritabilitas dan heterosis karakter penting hasil biji pada galur-galur DMR dan QPM telah dilaksanakan. Empat galur resisten pathogen bulai (DMR), yaitu Nei 9008, P 345, Ki 3, dan MR 10; tiga galur berkualitas protein tinggi (qpm), yaitu CML 161, CML 163, dan CML 172; dan hibrida silang tunggalnya yang dibentuk melalui rancangan persilangan line tester digunakan sebagai materi penelitian. Karakterkarakter sebagai berikut, termasuk tinggi tongkol pertama, jumlah biji per baris, berat biji per tongkol, dan hasil biji per plot memperlihatkan variabilitas yang luas. Heritabilitas

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dalam arti sempit (hns) karakter hasil biji, dan komponen hasilnya berkisar dari 0.01 sampai dengan 0.71. Heritabilitas dalam arti luas (hbs) berkisar dari 0.20 sampai dengan 0.74. Dari penelitian ini ditemukan fakta bahwa hibrida yang berasal dari persilangan antara galur yang memiliki latar belakang genetik yang jauh memperlihatkan heterosis yang tinggi.

2002; Kasim et al., 2003). Furthermore, Ruswandi et al. (2004) reported some QPM lines possessing high content of lysine dan tryptophan adapted to agroecosystem of Indonesia. Those QPM lines are CML 161, CML 163, CML 164, CML 165, dan CML 172 (Ruswandi et al., 2004). Introgressions of those important traits from introduced lines into local cultivars need accurate genetic information on heritability, variability, and heterosis of grain yield and resistance against the pathogen. Ruswandi et al. (2003) reported that the selected DMR and QPM lines had different genetic background as revealed by SSR markers. Hallauer and Miranda (1981) summarized that heritability for yield ranged from 13.8 to 24.4. The estimates of heritability for kernel row number, kernel weight (g), and cob diameter were 57.0; 41.8; 37.0; respectively. Martin and Hallauer (1976) as cited by Hallauer and Miranda (1981) reported that estimation of heterosis for yield, ear length, ear diameter, kernel row number, and 300 kernel weight ranged from 124.82 to 14.4%, 23.9% to 51.2%; 14.4% to 24.0%; 9.1% to 17.9%, and 4.0% to 19.6%; respectively. The objectives of this experiment were to study genetic variability and to estimate both heritability and heterosis of important grain yield on DMR and QPM lines and their single cross hybrids.

Introduction
Downy mildew (DM) which is caused by fungal from genus Perenosclerospora is the most devastating problem of maize production in Indonesia and other Asian countries including: India, Philippines, and Thailand (Azrai, et al., 2002; Ruswandi et al., 2002). Ideally, improvement of high yield maize cultivars possessing high quality of seed as shown by high lysine and tryptophan content must incorporate genes resistance to DM pathogen in any maize breeding program (Ruswandi, et al., 2004). Experiences in evaluating introduced materials under DM stress nursery in Maros showed high to very high susceptibility of QPM to the pathogen. High lysine and tryptophan maize is known as quality protein maize (QPM) due to opaque-2 gene. An integrated research on improvement of maize cultivars possessing those important traits has been starting since 1998 in Asia through Asian Maize Biotechnology Network (AMBIONET). As a result, some inbred lines were successfully selected as new sources of resistance against either P. maydis (the causal agent of downy mildew in Indonesia and Thailand) or P. philippinensis (the causal agent of downy mildew in Philippines and South Sulawesi province of Indonesia). Those new selected resistance lines are Ki3, AMATLCOHS, Nei 9008, and P 345 (Ruswandi, et al., 2002; Kasim et al.,

Materials and Methods

Four downy mildew resistance (DMR) lines, i.e. Nei 9008, P 345, Ki 3, and MR 10; three QPM lines, i.e. CML 161, CML 163, and CML 172; were used to develop single cross hybrids based on line tester mating design (Singh and Chaudhary, 1979). The single cross hybrids with enough seeds were as follow: Nei 9008 CML 161, Nei 9008 CML 163, Nei 9008 CML 172, P

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345 CML 161, P 345 CML 163, P 345 CML 172, Ki 3 CML 161, Ki 3 CML 163, Ki 3 CML 172, MR 10 CML 161, MR 10 CML 163, dan MR 10 CML 172. Those genotypes were planted in the field of Experimental Station, College of Agriculture, Padjadjaran University, Jatinangor-Bandung, Indonesia. The experiment was arranged in a randomized block design replicated twice. Each entry was grown in 5 m long single row plot at 0.75 between rows and 0.25 m within rows. The characters measured, included: height of first ear, number of ear, ear length, rows number per ear, seeds number per row, seed diameter, seed height, 100 seed weight, seed weight per ear, seed weight per plant, and grain yield per plot. Analysis of variance for each character was performed following calculation suggested by Singh and Chaudhary (1979) for line tester analysis. Genetic components such as additive variance, dominance variance, genetic variability, and heritability were estimated based on the ANOVA. The following formula suggested by Singh and Chaudhary (1979) were used in the calculation of the genetic component for each character: 2A = 4 2gca 2D = 4 2sca Broad sense heritability (h2bs) =
2 2 2

Mid parent (MP) and high parent (HP) heterosis were estimated following formula suggested by Allard (1960).

Results and Discussion

Genetic variability for grain yield of maize and its components were presented in Table 1. Of eleven characters, four characters, i.e., height of first ear, seed number per row, seed weight per ear, and grain yield per plot showed broad genetic variability. This result was in line with the earlier study by Ruswandi et al. (2004) who classified DMR and QPM parental lines into three groups that were genetically different based on SSR analysis. Furthermore they explained that SSR markers to be used were highly discriminative to determine DNA of inbred lines since it had high polymorphic index coefficient (PIC). Narrow sense heritability ( h 2 ns ) of grain yield and its components ranged from 0.01 to 0.71 (Table 1). None showed high narrow sense heritability. This result was similar to the extensive literature studies done by Hallauer and Miranda (1982) on estimation of narrow sense heritability based on different mating design (NC I, NC II, NC III, diallel, and generation mean analysis) using different population (F2, synthetics, open pollinated, variety crosses, and composites). They classified narrow sense heritability of grain yield and its components into low up to medium. Broad sense heritability ( h 2 bs ), on the other hand, ranged from 0.20 to 0.74 (Table 1). From the estimation of heritability, it is clear that narrow sense heritability had lower value than broad sense heritability because it estimates genetic variance of additive (Table 2). Non additive variance, i.e. dominance and epistacy variance, were not included in the estimation of narrow sense

D 2 f

Narrow sense heritability (h2ns) =

2A 2f

Genetic variability was estimated based on coefficient of variance genetics (CVG) following Burton et al. (1952).

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heritability since in most cases the component of genetic variability transmitted from parent to offspring is the additive component and other components will depend on the individuals or populations own combination of genes and not on its parents (Carpena, et al., 1993). The range of MP and HP heterosis for

heterosis; while Ki 3 CML161 and MR 10 CML 172 expressed the highest heterosis (Table 3). MP and HP heterosis for number of ear ranged from 37.04 to 10.51 and 41.38 to 10.51, respectively. The lowest heterosis was showed by Ki3 CML163. In contrast, Nei 9008 CML172 expressed the highest heterosis. Ear length possessed MP and HP heterosis that ranging from 5.33 to 50.06 and 5.10 to 43.18, respectively. P 345

height of first ear were from 4.03 to

78.65 and 20.23 to 68.29, respectively. P 345 CML172 showed the lowest

Table 1. Heritability and variability of grain yield and its components

h2 bs
No. 1 2 3 4 5 6 7 8 9 10 11 Characters Height of first ear Number of Ear Ear length Rows number per ear Seeds number per row Seed diameter Seed height 100 seed weight Seed weight per ear Seed weight per plant Grain yield per plot Value 0.20 0.66 0.63 0.58 0.68 0.74 0.68 0.69 0.61 0.64 0.61 Criteria Low high high high high high high high high high high Value 0.09 0.13 0.15 0.12 0.11 0.71 0.15 0.32 0.01 0.05 0.01

h2 ns
Criteria Low Low Low Low Low High Low Medium Low Low Low

Genetic Variability Genetic Variance CVG Criteria 227.378 0.001 1.907 0.116 26.929 0.002 0.005 5.029 440.344 18.539 176137.868 15.59 2.04 8.95 2.56 19.42 5.64 7.56 8.07 20.06 3.76 20.06 Broad Narrow Narrow Narrow Broad Narrow Narrow Narrow Broad Narrow Broad

Table 2. Genetic components of grain yield and its components

No. 1 2 3 4 5 6 7 8 9 10 11 Characters Height of first ear Number of ear Ear length Rows number per ear Seeds number per row Seed diameter Seed height 100 seed weight Seed weight per ear Seed weight per plant Grain yield per plot

2A
51.9939 1.3735 0.2550 0.6763 7.1686 0.0041 0.0034 18.7367 27.4576 249.9857 10983.0581

2D
58.8942 4.2995 9.1053 2.5594 35.6541 0.0002 0.0117 21.8246 1453.2994 2862.6761 581319.7666

2E
443.6612 3.2792 5.1734 2.3786 20.3452 0.0015 0.0070 18.0711 952.1602 1731.5650 380864.0708

2P
554.5493 8.9522 14.5337 5.6144 63.1679 0.0057 0.0222 58.6324 2432.9172 4844.2268 973166.8954

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CML 172 conveys the lowest MP, while Ki 3 CML 161 stated the lowest HP. On the contrary, MR 10 CML163 expressed the highest heterosis. The range of MP and HP heterosis for rows number per ear were from 14.42 to 31.91 and 16.96 to 22.82, respectively. MR 10 CML161 showed the lowest heterosis. In contrast, P345 CML163 expressed the highest heterosis. MP and HP heterosis for seeds number per row ranged from 29.05 to 104.89 and 15.76 to 80.53, respectively. The lowest heterosis was showed by Ki3

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Table 3. Heterosis of grain yield and its components on single cross hybrids of downy mildew resistance quality protein maize
HETEROSIS (%) Height of first ear MPH HPH 2 Ki3 CML163 MPH HPH 3 Ki3 CML172 MPH HPH 4 MR10 CML161 MPH HPH 5 MR10 CML163 MPH HPH 6 MR10 CML172 MPH HPH 7 Nei9008 CML161 MPH HPH 8 Nei9008 CML163 MPH HPH 9 Nei9008 CML172 MPH HPH 10 P345 CML161 MPH HPH 11 P-345 CML-163 MPH HPH 12 P345 CML172 MPH HPH 78.65 50.88 26.69 5.34 40.60 26.53 32.46 26.16 33.41 10.02 72.42 68.29 49.34 44.31 20.98 7.02 11.66 0.70 17.12 3.58 21.53 17.47 4.03 20.23 Number of ear 29.96 31.82 37.04 41.38 13.64 24.00 1.27 9.09 23.99 34.47 15.01 19.06 9.24 6.07 8.33 24.13 10.51 10.51 12.69 16.66 28.29 34.47 16.28 25.00 Ear length 8.14 5.10 28.95 15.43 12.59 9.61 33.45 28.23 50.06 43.18 24.75 19.68 29.58 24.29 40.49 34.30 20.71 15.60 19.59 19.36 16.98 7.28 5.33 5.31 Rows number per ear 12.70 13.76 7.77 2.98 4.11 2.62 14.42 16.96 8.60 0.62 3.69 4.04 5.52 1.28 1.30 6.40 0.38 4.41 7.25 2.21 31.91 22.82 3.76 5.88 Seeds number per row 29.05 15.76 65.95 28.19 43.51 31.13 47.56 35.32 79.04 63.18 51.59 36.45 69.14 60.73 104.89 80.53 62.59 51.53 34.14 21.31 70.31 32.43 36.77 26.03 Seed diameter 8.82 8.55 9.18 6.38 5.54 3.36 16.93 9.64 8.74 4.28 13.25 8.07 5.40 3.27 3.73 3.40 11.76 11.54 2.80 0.78 0.47 0.74 6.66 4.83 Seed height 8.34 7.20 18.35 10.79 12.60 3.69 25.20 21.77 25.84 22.24 24.19 18.58 17.79 11.59 22.73 22.48 20.37 18.02 18.41 17.45 20.93 13.48 13.63 4.88 100 seed weight 19.26 13.51 21.13 14.81 12.83 12.42 28.33 14.98 15.15 3.58 23.89 5.94 0.20 5.67 1.91 3.28 13.06 1.67 19.90 18.42 9.33 7.50 2.66 1.46 Seed weight per ear 43.39 40.21 91.82 53.11 63.94 47.59 92.40 78.89 128.46 97.17 87.61 85.23 93.04 69.31 131.96 111.96 96.66 86.85 64.33 58.44 112.27 67.63 33.83 18.96 Seed weight per plant 28.78 24.91 92.00 42.64 21.94 15.57 54.83 35.21 112.94 71.58 34.02 14.68 70.92 47.24 156.70 109.41 21.81 2.86 61.52 45.78 107.37 62.49 4.45 7.72 Grain yield per plot 43.39 40.21 91.82 53.11 63.94 47.59 92.40 78.89 128.46 97.17 87.61 85.23 93.04 69.31 131.96 111.96 96.66 86.85 64.33 58.44 112.27 67.63 33.83 18.96

#
1

SINGLE CROSS HYBRIDS Ki3 CML161

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CML161. The Nei 9008 CML163 hybrid, quite the opposite, expressed the highest heterosis. Seed diameter express MP and HP heterosis that ranging from 0.47 to 16.93 and 0.78 to 9.64, respectively. P345 CML163 possessed the lowest MP heterosis; as P345 CML161 stated the lowest HP heterosis. In contrast, MR 10 CML161 conveyed the highest heterosis for this character. The range of MP and HP heterosis for seed height were from 8.34 to 25.84 and 3.69 to 22.24, respectively. Ki3 CML 161 showed the lowest MP heterosis, whereas Ki 3 CML 172 stated the lowest HP heterosis. The MR 10 CML163 cross, on the other hand, expressed the highest heterosis. MP and HP heterosis for 100 seed weight ranged from 0.20 to 28.33 and 5.67 to 18.42, respectively. The lowest heterosis was showed by Nei 9008 CML161. Quite the reverse, MR 10 CML161 and P 345 CML 161 conveyed the highest heterosis. MP and HP heterosis for seeds weight per ear ranged from 33.83 to 131.96 and 18.96 to 111.96, respectively. The lowest heterosis was showed by P 345 CML 172. The Nei 9008 CML163 hybrid, quite the opposite, stated the highest heterosis. The range of MP and HP heterosis for seed weight per plant were from 4.45 to 156.70 and 7.72 to 109.41, respectively. P 345 CML172 showed the lowest heterosis. On the other hand, the Nei 9008 CML 163 hybrid expressed the highest heterosis. Grain yield per plot possessed MP and HP heterosis that ranging from 33.83 to 131.96 and 18.96 to 111.96 respectively. P 345 CML172 conveyed the lowest heterosis. In contrast, Nei9008 CML163 expressed the highest heterosis for this character.

It is found that hybrid derived from crossing between parental inbred lines possessing broad genetic background expressed high heterosis. The genetic background of Nei 9008, MR 10, and CML 163 are broad since they are classified in different genetic group based on cluster analysis. In addition, they are highly polymorphic as shown by high polymorphic index coefficient (PIC), for example single cross hybrid of MR 10 CML 163 showed high heterosis for five characters since their parental lines classified into different genetic group where CML 163 is in group 1, while MR 10 was in group 3 (Ruswandi, et al., 2003). Furthermore, MR 10 and CML 163 were highly polymorphic as showed by their PIC at 75% (Ruswandi, et al., 2004). In contrast, Ki 3 CML 161 had low heterosis since their parental lines classified in the same genetic group, that was group one (Ruswandi, et al., 2003). In addition, Ki 3 and CML 161 were low polymorphic as showed by their PIC at less than 50% (Ruswandi, et al., 2004).

Conclusions
Height of first ear, seed number per row, seed weight per ear, and grain yield per plot showed broad genetic variability. Narrow sense heritabilities (hns) of grain yield and its components ranged from 0.01 to 0.71. Broad sense heritability (hbs), on the other hand, ranged from 0.20 to 0.74. It is found that hybrid derived from crossing between parental inbred lines possessing broad genetic background expressed high heterosis.

Acknowledgment
The authors would like to put into words their appreciation to the

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following agencies: The Asian Maize Biotechnology Network (AMBIONET) for genetic materials; Ministry of Research and Technology, Republic of Indonesia for research funding through Riset Unggulan Terpadu granted to 1st author.

2003. Marker assisted for downy mildew resistance and diversity study of Indonesian lines. Unpublished. Ambionet Indonesia Semi Annual Progress Report. April 2003September 2003. Ambionet Annual Meeting. 36 November 2003. Chiang Mai. Thailand. Ruswandi, D., D.M. Hautea, A.L. Carpena, R.M. Lantican, A.M. Salazar, and A.D. Raymundo. 2002, Quantitative trait loci mapping of Philippines downy mildew resistance gene in maize (Zea mays L.). Zuriat. Indonesian Journal of Breeding Vol. 13. No. 1: 2734. Ruswandi, D., N. Wicaksana, M.B. Pabendon, M. Azrai, F. Kasim, M. Rachmady, A. Ismail, F. Damayanti, Nono Carsono, and T. Gunawan. 2003. Genetic relationship of quality protein of maize and downy mildew resistance lines based on simple sequence repeats (SSR) marker. International Seminar on Biotechnology for Sustainable Agriculture: State of the Art of Research and Product Commercialization. October 78, 2003. SEAMEO BIOTROP. Bogor. Indonesia. Ruswandi, D., N. Carsono, N. Wicaksana, M.B. Pabendon, M. Azrai, F. Kasim, M. Rachmadi, and A. Ismail. 2004. DNAfingerprinting of downy mildew resistance (DMR) and quality protein maize (QPM) lines using eleven simple sequence repeats (SSR). In: Proceedings of Low Input Sustainable Agriculture I. D. Ruswandi, and N. Carsono (Eds.). Department of Crop Science, Faculty of Agriculture, Padjadjaran University, Bandung, Indonesia. p. 5666. Singh, H.K., and B.D. Chaudhary. 1979. Biometrical Methods in Quantitative Genetic Analysis. Kalyani Publisher. Ludhiana. New Delhi. p. 191200.

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