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Gn1a and TGW6 Targeted Knock- Down via Genome editing Increases Yield Components

in Samba Mahsuri.

Lawrence Uy1,2, Yvonne Ludwig2 and Inez Slamet-Loedin2

1
University of the Philippines-Los baños, 2International Rice Research Institute

ABSTRACT

Rice shortage is one of the world’s biggest problems. An estimate of 3 billion people

depends on it. One approach in tackling this problem is through genome engineering of yield-

related genes of elite rice varieties. The genes THOUSAND GRAIN WEIGHT 6 (TGW6) and

GRAIN NUMBER 1A (Gn1a) are known to influence the grain weight and number of

reproductive organs, respectively. In this study, TGW6 and Gn1a were specifically targeted in

Samba Mahsuri (SM) using Clustered Regularly Interspaced Short Palindromic Repeats

(CRISPR)-Cpf1 system. Two constructs, IRS1466 targeting TWG6 and IRS1467 targeting Gn1a,

were used to generate SM transgenic plants. Transgenic Samba Mahsuri lines were analyzed for

the targeted variation in the promoter region or gene of interest. The analysis revealed that the

sgRNA efficiency significantly differed between the constructs. SM-IRS1466 had 94.34% while

SM-IRS1467 displayed 21.23% efficiency. Phenotypic data showed a significant increase in

thousand grain weight and total grain weight of SM-IRS1466 transgenic lines compared to the

wild-type control. The thousand grain weights of SM-IRS1466-285 increased by 26% while the

total grain weight of SM-IRS1466-26 increased by 33.5%. SM-IRS1467 lines showed significant

increase in total grain weight. SM-IRS1467-169 gained 80% more total grain weight. These

results reveal that targeted knockdown of TGW6 and Gn1a can be used in increasing yield in the

elite rice variety Samba Mahsuri.