BIODIVERSITAS ISSN: 1412-033X Volume 22, Number 12, December 2021 E-ISSN: 2085-4722 Pages: 5425- 5431 DOI: 10.13057/biodiv/d221225 Selection of doubled haploid black rice lines in advanced yield trial based on multivariate analysis ALIM WULANDARI KARIMA 1, , RATNA KARTIKA PUTRI 2 , BAMBANG SAPTA PURWOKO 2, , ISWARI SARASWATI DEWI 3 , WILLY BAYUARDI SUWARNO 2 , ANI KURNIAWATI 2 1 Graduate School, Institut Pertanian Bogor. Jl. Raya Dramaga, Kampus IPB, Bogor 11680, West Java, Indonesia.  email: wdari1500@gmail.com 2 Department of Agronomy and Horticulture, Faculty of Agriculture, Institut Pertanian Bogor. Jl. Meranti, Kampus IPB Dramaga, Bogor 16680, West Java, Indonesia. Tel./fax: +62-251-8629353,  email: bspurwoko@apps.ipb.ac.id 3 Indonesian Center for Agricultural Biotechnology and Genetic Resources Research and Development. Jl. Tentara Pelajar No. 3A, Bogor 16111, West Java, Indonesia Manuscript received: 26 August 2021. Revision accepted: 22 November 2021. Abstract. Karima AW, Putri RK, Purwoko BS, Dewi IS, Suwarno WB, Kurniawati A. 2021. Selection of doubled haploid black rice lines in advanced yield trial based on multivariate analysis. Biodiversitas 22: 5425- 5431. Research on the development of black rice has not received much attention. The anther culture technique can shorten the plant breeding process and produce doubled haploid (DH) lines. The target of this study was to evaluate DH black rice lines with early maturity, good agronomic performance, and high yields. Twenty- seven doubled haploid lines and three check varieties (Aek Sibundong, Inpari 24, and Jeliteng) were evaluated at two locations (Kediri, East Java, and Bogor, West Java) from January to July 2020. A randomized complete block design was applied to this study with three replicates. The combined analysis results showed that the genotype effect (G) was highly significant (p < 0.01) for all characters. In addition, all characters had a high broad-sense heritability. Characters that were positively correlated with yield based on genotypic and phenotypic correlation were the number of productive tillers, the number of vegetative tillers, panicle length, the number of filled grains, and the number of total grains. The 1000-grains weight and panicle length had a positive and the largest direct effect on yield. Principal component analysis showed that the number of productive tillers and the 1000-grains weight were related to yield. The selection index model that can be formulated is as follows I = (3*0.375*Yield) + (0.723* 1000-grains weight) + (0.280* Number of productive tillers). Based on the selection index, 14 genotypes were selected. Keywords: Anther culture, combined analysis of variance, correlation, functional food path analysis, principal component analysis INTRODUCTION Black rice, included in the Oryza sativa L., can be used as a functional food and has health-related benefits (Pratiwi and Purwestri 2017). It is because of anthocyanin content, phenolic acid, flavonoids, and antioxidant activity, which positively correlated to each other and higher than that of red and white rice (Pengkumsri et al. 2015). A study reported that the anthocyanin content in black rice is 5.121 ± 0.521 mg GAE/g, flavonoid content is 9.824 ± 1.546 mg QE/g and antioxidant capacity is 6.308 ± 0.318 mg AEAC/g (Yuliana and Akhbar 2020). The anthocyanin is the expression of the Kala4 gene and accumulates in the pericarp (Oikawa et al. 2015). The anthocyanin can be antihyperlipidemic, antihyperglycemic, to prevent hyper diabetic syndrome in fructose-fed, and used as dietary supplements (Prasad et al. 2019). The anthocyanin in black rice has a function as an antidote to free radicals and an antioxidant. The antioxidant activity of black rice is directly proportional to its phenolic, flavonoid, and anthocyanin content (Phan et al. 2018; Priya et al. 2019). Research on the development of black rice has not received much attention due to the limited varieties available for planting seeds, late maturity, low yield, and susceptibility to bird pests (Hanifah et al. 2016). One approach to solve this problem involves the use of biotechnology breeding techniques such as anther culture to develop black rice varieties with early maturity and high-yielding ability. The anther culture technique supported doubled haploid (DH) homozygous plants production from the F1. Plant breeding process with anther culture technique is shorter than other conventional rice plant breeding methods which take very long time (7-10 years) to get pure lines (Dewi and Purwoko 2012). Previous researches successfully produced fertile DH lines that have good agronomic performances. Azmi et al. (2017) produced DH lines from anther culture of the F1 produced from crosses between lowland and cultivated black rice varieties. Maharani et al. (2020) induced and regenerated callus for black rice indica, while Mawaddah et al. (2018) produced the DH lines of red rice with high yield from anther culture. Alsabah et al. (2019) characterized DH lines in the preliminary yield test (PYT). DH lines must be evaluated further in the advanced yield test (AYT). AYT is one of the steps needed to test lines before being selected and tested in several locations to be re-selected again as potential candidates of varieties (Endelman et al. 2014). The selection used in this research was index selection. Index selection uses several characters (secondary characters) simultaneously in a linear model. Index selection aims to increase the genotypic value in the population (Hazel et al. 1994; Sabouri et al. 2008).