Simple-sequence-repeat (SSR) and PCR-RFLP were employed to characterize the nuclear and cytoplasmic genomes of intergeneric diploid plants derived from symmetric fusion between Microcitrus papuana Swingle and Rough lemon (Citrus jambhiri Lush). Three out of five SSR primers distinguished the fusion parents from each other and the regenerated plants showed band profiles completely identical to the leaf parent, Rough lemon. Amplified products from the intergenic regions of cpDNA between trnD-trnT were digested with HaeIII and MspI, and those between trnH-trnK were digested with HinfI, and both the regenerated plants and Rough lemon shared the same band patterns, which were different from the embryogenic parent, M. papuana. With mtDNA, only 2 out of 12 primer pair/restriction enzyme combinations (nad4 ex 1-2/TaqI and nad4 ex 1-2/HindIII) revealed polymorphisms between the fusion parents. With the former combination the regenerated plants showed the same fragment distribution as that of the embryogenic parent, M. papuana, whereas with the latter, a novel band absent in the fusion parents was detected in all of the regenerated plants, suggesting a possible rearrangement. The present research indicates that the plants analyzed were putative cybrids containing nuclear DNA and cpDNA from Rough lemon and mtDNA from M. papuana. Presumed mechanisms leading to the regeneration of diploid hybrid plants following symmetric fusion are discussed herein.

Polyamine uptake transporter (PUT) plays important roles in polyamine homeostasis, but knowledge regarding PUT family genes in sweet orange (Citrus sinensis) remains elusive.• Herein, our study aimed to perform a genome-wide identification of the PUT gene family in C. sinensis. A total of eight putative PUT genes (CsPUT1-CsPUT8) were identified in the sweet orange genome and distributed on three chromosomes. The CsPUT genes were divided into two major groups according to the phylogenetic tree analysis, with high similarities in protein domains and gene structure organization.• The CsPUT genes were differentially expressed in different tissues, with the highest transcript levels being in the flowers and roots. Interestingly, the CsPUT genes were significantly induced by polyamines, putrescine, spermidine and spermine, indicating that CsPUT were possibly associated with intracellular polyamine transport and uptake. In addition, CsPUT showed differential expression in callus treated with ABA, cold, salt or osmotic shock. • CsPUT4 was selected as a candidate for functional analysis of PUT. Overexpression of CsPUT4 elevated endogenous polyamine content and led to enhanced cold tolerance in transgenic callus cultures. Overall, these data provide valuable information for better understanding the potential biological functions of PUT genes in future.