Chuchu Liu^1,2 , Zhonggang Li¹

1 Cuixi Academy of Biotechnology, Zhuji, 311800, Zhejiang, China
2 Hainan Institute of Tropical Agricultural Resources, Sanya, 572025, Hainan, China
Author    Correspondence author
Bioscience Methods, 2026, Vol. 17, No. 1   
Received: 01 Dec., 2025    Accepted: 04 Jan., 2026    Published: 19 Jan., 2026

This is an open access article published under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

The three main causes of yield loss in pineapple crops include heart rot and black rot and leaf spot. Breeding new types in the usual way takes many years. The crop maintains a restricted genetic diversity because it cannot produce self-pollination and requires extended periods for growth and testing. The research evaluates CRISPR/Cas9 as a fast method to introduce disease resistance. We show the main defense routes, such as SA, JA/ET, and MAPK. We implement editing approaches which have proven effective in different plant species. The main ways are: (i) change promoters or switch on defense genes; (ii) remove susceptibility genes and genes that decrease defense; (iii) modify control sites such as miRNA binding sites; and (iv) edit multiple genes simultaneously to build resistance. Some problems remain. The regeneration rate is low and edited plants can be mosaic and pineapple has high heterozygosity and off-target hits can occur. The solutions for this problem include tissue culture improvement and morphogenic regulator addition and nuclease precision enhancement and RNP delivery without DNA. Our approach consists of three phases which begin with omics-based target selection followed by DNA-free multiplex editing and end with field testing and compliance with transgene-free plant regulations. The described methods enable pineapples to develop robust and extensive resistance that can be used by breeders to create new plant lines. Keywords Pineapple; CRISPR/Cas9; Disease resistance; Susceptibility genes; Promoter editing; Multiplex editing; DNA-free delivery

Pineapple (Ananas comosus); CRISPR/Cas9; Disease resistance; Susceptibility genes; Promoter editing; Multiplex editing; DNA-free delivery