Chlorosis resistant cytoplasmic male sterile Brassica plants

A method of producing a chlorosis resistant CMS Brassica rapa plant, comprising the steps of:

• a) protoplast fusion of protoplasts of a fertile Brassica rapa plant with protoplasts of a CMS Brassica rapa plant as a CMS cytoplasm donor, wherein the cytoplasm donor comprises chloroplasts of a diploid Brassica oleracea plant and Raphanus sativus mitochondria, wherein mitochondria of the CMS cytoplasm donor are combined with chloroplasts and nucleus of the fertile Brassica rapa plant;
• and
• b) selecting a protoplast fusion product, wherein:
• i. at least 95% of fusion product chloroplasts are Brassica rapa chloroplasts comprising one or more sequences selected from the group consisting of SEQ ID NO:
• 1, SEQ ID NO:
• 3 and SEQ ID NO:
• 5;
• ii. the fusion product nuclear genome is a Brassica rapa nuclear genome comprising SEQ ID NO:
• 7 and/or SEQ ID NO:
• 9;
• iii. the fusion product does not comprise Brassica oleracea chloroplasts that comprise one or more sequences selected from the group consisting of SEQ ID NO:
• 2, SEQ ID NO:
• 4, and SEQ ID NO:
• 6;
• and
• iv. the fusion product comprises a mitochondrial genome of Raphanus sativus comprising the sequence of SEQ ID NO:
• 11.