Ciacculli, A.

Larger, S.

Licciardello, C.

Malnoy, M.

Pappalardo, H. D.

Pindo, M.

Poles, L.

Salonia, F.

English

Archivio istituzionale della ricerca - Fondazione Edmund Mach

1/31/23, 9:24 AM Plant and Animal Genome 30https://plan.core-apps.com/pag_2023/abstract/cc642af8-8869-4767-b129-7955c8139ad8 1/2The Use of Modern Biotechnology Approaches to ImproveCitrus Fruit QualityAuthorsAngelo Ciacciulli (/pag_2023/speaker/dd20ad965c578f0cb65c90015f89d699)Fabrizio Salonia (/pag_2023/speaker/4ac8a4037afdedea5bee9d19080b5cc4)Lara Poles (/pag_2023/speaker/b3177b1ca1cd14df757784d22cfbf76a)Helena Domenica Pappalardo (/pag_2023/speaker/b3177b1ca1cd14df757784d22cfbf61e)Simone Larger (/pag_2023/speaker/dd20ad965c578f0cb65c90015f89ce0c)Massimo Pindo (/pag_2023/speaker/dd20ad965c578f0cb65c90015f89dfa9)Mickael Malnoy (/pag_2023/speaker/dd20ad965c578f0cb65c90015f89bfdb)Concetta Licciardello (/pag_2023/speaker/e18f133d173d03ad9aa8173bb9c28638)BodyAdditional Categories: Plants: Trees|forestry, Technology: Sequencing, Plants and Animals: Others, such as Functional Genomics, Plants and Animals:Population, Conservation and Ecological GenomicsFruit species are suffering of the difficulties related to classical breeding techniques, in terms of financial commitment, land resources availability and longgeneration times. The most recent ‘new genomic techniques’ (NGTs) reduce the time required to obtain new varieties as well as to precisely target specificDNA sequences without altering any other trait.We used the NGTs to improve two important traits for consumer needs consisting in the combination of anthocyanins and lycopene in a unique fruit, and inreducing seeds content in mandarin and mandarin-like varieties. For the first aim, we introduced Ruby (the Myb-like transcription factor known for regulatingthe anthocyanin synthesis in citrus fruits) as a cisgene in lycopene-rich accessions, using a vector containing a FLP/FRT system to obtaining marker-freecisgenic plants. We are optimizing the heat shock treatment so to excise the entire cassette containing the kanamycin resistance. On the other hand, weperformed the knockout of β-LCY2, through a dual-single guide (sgRNA) approach using the GoldenBraid 3.0 technology, to produce loss-of-function mutantsable to induce lycopene accumulation in anthocyanin-rich sweet oranges. The amplicons sequencing and the CRISPResso tool confirmed the presence ofseveral editing events.On the second aim, seedlessness in citrus is a complex trait under the control of several biological processes. So far, any candidate genes that can bemanaged to produce seedless varieties. Therefore, we decided to edit Iku1, the homologous of Arabidopsis, which loss of function mutations cause adecrease in seed size. Two sgRNAs were designed to knock-out Iku1 in ‘Carrizo’ citrange and in sweet orange. Several plantlets with different editing eventshave been produced.With the aim to speed-up the fruits production so to see the effects of the cisgenic and genome editing strategies we approached, we also produced a vectorcontaining the Cas9 and the sgRNA in order to induce the knockout of TFL1, the gene responsible of the precocious flowering. Our final purpose is toproducing, in the next future, early flowering citrus plants producing highly healthy fruits.Our work represents the unique examples in which NGTs, so far, were used to improve the citrus fruits quality.SessionsThe Use of Modern Biotechnology Approaches to Improve Citrus Fruit QualitySaturday, Jan 14 10:19 AMPacific E1/31/23, 9:24 AM Plant and Animal Genome 30https://plan.core-apps.com/pag_2023/abstract/cc642af8-8869-4767-b129-7955c8139ad8 2/2(/pag_2023/event/85ddbebe25a2e87ae2f282b9e9d6715c)

The use of modern biotechnology approaches to improve citrus fruit quality

https://openpub.fmach.it/bitstream/10449/78237/1/Plant%20and%20Animal%20Genome%2030%20citrus.pdf

The use of modern biotechnology approaches to improve citrus fruit quality

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Archivio istituzionale della ricerca - Fondazione Edmund Mach