The jujube witches’ broom (JWB) disease is a severe threat to jujube trees, with only a few cultivars being genuinely tolerant or resistant to phytoplasma. The defense mechanism of jujube trees against phytoplasma is still unclear. In this study, we aimed to investigate the tolerance mechanism of Indian jujube ‘Cuimi’ to JWB and identify the key genes that contribute to JWB high tolerance. Based on the symptoms and phytoplasma concentrations after infection, we confirmed the high tolerance of ‘Cuimi’ to JWB. Comparative transcriptome analysis was subsequently performed between ‘Cuimi’ and ‘Huping’, a susceptible cultivar of Chinese jujube. Unique gene ontology (GO) terms were identified in ‘Cuimi’, such as protein ubiquitination, cell wall biogenesis, cell surface receptor signaling pathway, oxylipin biosynthetic process, and transcription factor activity. These terms may relate to the normal development and growth of ‘Cuimi’ under phytoplasma infection. We identified 194 differential expressed genes related to JWB high tolerance, involved in various processes, such as reactive oxygen species (ROS), Ca²⁺ sensors, protein kinases, transcription factors (TFs), lignin, and hormones. Calmodulin-like (CML) genes were significantly down-regulated in infected ‘Cuimi’. We speculated that the CML gene may act as a negative regulatory factor related to JWB high tolerance. Additionally, the cinnamoyl-CoA reductase-like SNL6 gene was significantly up-regulated in infected ‘Cuimi’, which may cause lignin deposition, limit the growth of phytoplasma, and mediate immune response of ‘Cuimi’ to phytoplasma. Overall, this study provides insights into the contribution of key genes to the high tolerance of JWB in Indian jujube ‘Cuimi’.

The Xyloglucan endotransglucosylase/hydrolase (XTH) family, a group of cell wall-modifying enzymes, plays crucial roles in plant growth, development, and stress adaptation. The quality and yield of Chinese jujube (Ziziphus jujuba) fruit are significantly impacted by environmental stresses, including excessive salinity, drought, freezing, and disease. However, there has been no report of the XTH encoding genes present in the Chinese jujube genome and their response transcription level under various stresses. This study provides an in-depth analysis of ZjXTH genes in the genome of Chinese jujube and elucidates their structural motifs, regulatory networks, and expression patterns under various stresses. A total of 29 ZjXTH genes were identified from the Ziziphus jujuba genome. Phylogenetic analysis classifies ZjXTH genes into four distinct groups, while conserved motifs and domain analyses reveal coordinated xyloglucan modifications, highlighting key shared motifs and domains. Interaction network predictions suggest that ZjXTHs may interact with proteins such as Expansin-B1 (EXPB1) and Pectin Methylesterase 22 (PME22). Additionally, cis-regulatory element analysis enhances our understanding of Chinese jujube plant’s defensive systems, where TCA- and TGACG-motifs process environmental cues and orchestrate stress responses. Expression profiling revealed that ZjXTH1 and ZjXTH5 were significantly upregulated under salt, drought, freezing, and phytoplasma infection, indicating their involvement in biotic and abiotic stress responses. Collectively, these findings deepen our understanding of the functional roles of Chinese jujube XTHs, emphasizing their regulatory function in adaptive responses in Chinese jujube plants.

The selection of unique and individual-specific SNPs is important as compared with universal SNPs for individual identification. Therefore, the main significance of this research is the selection of specific SNPs in male parent and the identification of offspring with these specific SNPs in their genome by multiplex PCR, which is utilized for genotyping of 332 half-sib plants of Ziziphus jujuba.This cost-effective method makes as much as possible to utilize the same amount of each pair of various targeted loci primers. After PCR amplification of targeted genome parts, the mixed products can be directly used in a next-generation sequencing platform. We concomitantly amplified 10 unique SNP loci at 10 different chromosomes of male JingZao 39 plants in 332 half-sib plants and sequenced them on the Illumina Novaseq 6000 platform. Analysis of SNP genotyping accuracy of 332 half-sib plants showed that all 10 unique SNPs in all 332 plants were correctly amplified in this multiplex PCR method. Furthermore, based on Mendelian inheritance, we identified 124 full-sib plants that have 10 unique SNPs in their genomes. These results were further confirmed by whole genome resequencing of 82 randomly selected half-sib plants, and the identity-by-descent values of all full-sib plants were between 0.4399 to 0.5652. This study displayed a cost-effective multiplex PCR method and proper identification of pollen parent through specific SNPs in half-sib progenies and firstly obtained a full-sib population between ‘Wuhezao’ and ‘JingZao 39’, segregating for stone and stoneless fruit.

The Xyloglucan endotransglucosylase/hydrolase (XTH) family, a group of cell wall-modifying enzymes, plays crucial roles in plant growth, development, and stress adaptation. The quality and yield of Chinese jujube (Ziziphus jujuba) fruit are significantly impacted by environmental stresses, including excessive salinity, drought, freezing, and disease. However, there has no report of the XTH encoding genes present in Chinese jujube genome and their response transcription level under various stresses. This study provides an in-depth analysis of ZjXTH genes in genome of Chinese jujube and elucidating their structural motifs, regulatory networks, and expression patterns under various stresses. A total of 29 ZjXTH genes were identified from Ziziphus jujuba genome. Phylogenetic analysis classifies ZjXTH genes into four distinct groups, while conserved motifs and domain analyses reveal coordinated xyloglucan modifications, highlighting key shared motifs and domains. Interaction network predictions suggest that ZjXTHs may interact with proteins such as Expansin-B1 (EXPB1) and Pectin Methylesterase 22 (PME22). Additionally, cis-regulatory element analysis enhances our understanding of Chinese jujube plant's defensive systems, where TCA- and TGACG-motifs process environmental cues and orchestrate stress responses. Expression profiling revealed differential regulation of ZjXTH genes under various stress conditions. Under salt stress, ZjXTH1 was upregulated, while ZjXTH6, and ZjXTH13 were downregulated, with ZjXTH3 and ZjXTH19 exhibiting negative regulation. In response to drought, ZjXTH5 showed upregulation, whereas ZjXTH7, -25, -13 were downregulated, and ZjXTH13 and ZjXTH19 were negatively regulated. Furthermore, under cold conditions, ZjXTH14 and ZjXTH18 were upregulated, while ZjXTH22, -23, -28, -29 were downregulated, and ZjXTH20 exhibited negative regulation. Additionally, phytoplasma infection induced the upregulation of ZjXTH1, -3, -5, with ZjXTH6, -7, -11, -16, -25 being downregulated, and ZjXTH20 showing negative regulation. Collectively, these findings deepen our understanding of the functional roles of Chinese jujube XTHs, emphasizing their regulatory function in adaptive responses in Chinese jujube plants.

An efficient callogenesis and regeneration protocol was optimized for six different wheat (Triticum aestivum L.) varieties including Chakwal-50, Galaxy-01, NARC-09, Pakistan-13, Millat-11, and Borlog-14. For callogenesis, mature seeds of each variety were used as explant, surface sterilization, and culturing were carried out on MS medium enhanced with dissimilar levels (0.5, 1.5, 2.5, 3.5, 4.5, and 5.5 mg/L) of 2,4-dichlorophenoxyacetic acid (2,4-D). For regeneration, six different treatments with different combinations of IAA (1.5, 3, and 4.5 mg/L) and Kinetin (1, 2, 3, and 4 mg/L) were applied on each variety. Highest frequency of callogenesis (41.66%) was attained on MS medium in addition with 2.5, 5.5 and 3.5 mg/L 2,4-D in Millet-11, followed by Chakwal-50 (33.33%) and Borlog-14 (29.16%), respectively. Maximum regeneration (41.66%) was observed in Millat-11 on medium enhanced with 1.5 mg/L IAA and 2 mg/L Kinetin. Among all, Millet-11 appeared most responsive genotype towards callogenesis and regeneration, while NARC-09 (16.66%) was least responsive. This study will open new avenues for biofortification of Mellet-11 wheat variety and improvement against biotic and abiotic stress. Key words: Wheat, tissue culture, callogenesis, regeneration, Millet-11, 2,4-D Kinetin. An efficient callogenesis and regeneration protocol was optimized for six different wheat (Triticum aestivum L.) varieties including Chakwal-50, Galaxy-01, NARC-09, Pakistan-13, Millat-11, and Borlog-14. For callogenesis, mature seeds of each variety were used as explant, surface sterilization, and culturing were carried out on MS medium enhanced with dissimilar levels (0.5, 1.5, 2.5, 3.5, 4.5, and 5.5 mg/L) of 2,4-dichlorophenoxyacetic acid (2,4-D). For regeneration, six different treatments with different combinations of IAA (1.5, 3, and 4.5 mg/L) and Kinetin (1, 2, 3, and 4 mg/L) were applied on each variety. Highest frequency of callogenesis (41.66%) was attained on MS medium in addition with 2.5, 5.5 and 3.5 mg/L 2,4-D in Millet-11, followed by Chakwal-50 (33.33%) and Borlog-14 (29.16%), respectively. Maximum regeneration (41.66%) was observed in Millat-11 on medium enhanced with 1.5 mg/L IAA and 2 mg/L Kinetin. Among all, Millet-11 appeared most responsive genotype towards callogenesis and regeneration, while NARC-09 (16.66%) was least responsive. This study will open new avenues for biofortification of Mellet-11 wheat variety and improvement against biotic and abiotic stress. Key words: Wheat, tissue culture, callogenesis, regeneration, Millet-11, 2,4-D Kinetin.