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The most common rootstock for Juglans regia (Persian or “English” walnut) in California is Paradox, typically a hybrid of J. hindsii (Northern California black walnut) × J. regia. Unfortunately, Paradox is very susceptible to Armillaria root disease. The relative resistance to Armillaria mellea of six clonally propagated Paradox rootstocks (AX1, Px1, RR4 11A, RX1, Vlach, VX211) was evaluated and compared with that of clonally propagated J. hindsii rootstock selection W17, J. regia scion cultivar Chandler, and Pterocarya stenoptera (Chinese wingnut). In a growth-chamber assay, plants were micropropagated and rooted in vitro before inoculating the culture medium with A. mellea. At two months post-inoculation, the most resistant and susceptible Paradox rootstocks were AX1 and VX211, respectively, with 9% vs. 70% mortality, and this finding was consistent across three isolates of A. mellea and three replicate experiments. This broad range of resistance within Paradox is consistent with past field trials that tested other genotypes. Our finding of similarly high susceptibility of ‘Chandler’ and W17 (61% vs. 69% mortality) is in contrast to two field trials, in which other J. regia genotypes were more susceptible than those of J. hindsii. A third trial, however, identified some J. regia genotypes as more resistant than those of J. hindsii. Therefore, it is possible that W17, which was not previously tested, is an Armillaria-susceptible genotype of J. hindsii. Based on our findings of repeatable mortality levels across three isolates of A. mellea and three replicate experiments, the growth-chamber assay has promise, albeit with confirmed resistant and susceptible controls, for identifying putative resistant rootstocks (e.g., AX1) in preparation for a field trial with controlled inoculations.

Prunus dulcis (almond) is one of the most susceptible horticultural crops to Armillaria root disease. Resistance to Armillaria mellea and Armillaria tabescens, the geographically isolated causal fungi that attack almond and closely related Prunus persica (peach), has been evaluated in studies of almond, peach, and other Prunus rootstocks, but not in one comprehensive study. We evaluated the relative resistance to A. mellea and A. tabescens of six clonally propagated almond and peach rootstocks (Bright’s 5, Empyrean 1, Hansen 536, Krymsk 1, Krymsk 86, and Lovell) in comparison with that of clonally propagated Marianna 2624 rootstock (resistant control) and clonally propagated Nemaguard rootstock (susceptible control). Replicate clones used in the growth chamber assay were micropropagated and rooted in vitro before inoculating the culture medium with Armillaria spp. At 2 months, the most resistant and susceptible rootstocks were Krymsk 86 and Hansen 536, respectively, with 27% vs. 89% mortality. This finding was consistent among two isolates of A. mellea and one isolate of A. tabescens in three replicate experiments. Our finding of low mortality among Krymsk 86, Krymsk 1, and Marianna 2624, which all share Prunus cerasifera (Myrobalan plum) parentage, is consistent with past reports of resistance in the field to A. mellea, but conflicts with reports of susceptibility to A. tabescens. Resistance to A. tabescens of genotypes with Myrobalan plum parentage in our assay may reflect the simplified rooting environment of tissue culture medium, which does not perfectly mimic a field trial, in which biotic and abiotic factors may affect host resistance. Nonetheless, our growth chamber assay may provide a more rapid alternative to identify sources of resistance for breeding and to screen progeny of such crosses.

Plants respond to pathogens with both active and passive defense mechanisms. These defense responses include the induction of defense or defense-related genes such as polyphenol oxidase (PPO) and pathogenesis-related (PR) proteins. The role of PPO in the interaction between bacterial blight [Xanthomonas arboricola pv. juglandis (Xaj)] and walnut (Juglans regia) was studied. JrPPO-1 and P14a genes were identified in two walnut cultivars, Chandler and Serr, using standard polymerase chain reaction (PCR) to understand their inducible ability in response to Xaj. ‘Serr’ and ‘Chandler’ were inoculated with Xaj strain 417. PPO activity in leaves was assayed at 0, 24, 72, 96, 120, and 144 hours after inoculation. Results showed a steady increase in activity commencing within 24 hours of inoculation. Increase in PPO activity was close to 2-fold greater in ‘Chandler’ than in ‘Serr’ at all time points examined. Real-time PCR analysis showed differences between cultivars in PPO gene expression. The JrPPO-1 gene was highly expressed in both cultivars 24 hours after inoculation but expression in ‘Serr’ was much greater than in ‘Chandler’. Significant expression of P14a gene was observed in both cultivars within 24 hours. Expression in ‘Serr’ was strong and maximized with a significant increase at 96 hours. Expression in ‘Chandler’ was far weaker than ‘Serr’ at 24 hours and did not increase further. Our results imply that the walnut–bacterial blight interaction induces the expression of JrPPO-1 and P14a as well as the activity of PPO.

Species of Phytophthora are serious soilborne pathogens of persian (english) walnut, causing crown and root rot and associated production losses worldwide. To facilitate the development of improved walnut rootstocks, we examined resistance of 48 diverse clones and seedlings of Juglans species to P. cinnamomi and P. citricola. Plants were micropropagated, acclimatized to a greenhouse environment, and then exposed to the pathogens in artificially infested potting soil mix. Inoculated plants, as well as noninoculated controls, were subjected to soil flooding for 48 hours every 2 weeks to facilitate infection by the pathogens. Two to 3 months after inoculation, resistance to the pathogens was assessed according to the severity of crown and root rot. Clonal hybrids of J. californica × J. regia were highly susceptible to the pathogens (means 52% to 76% root crown length rotted), while several clones of J. microcarpa × J. regia were significantly less susceptible (means 8% to 79% crown length rotted). Among clones of other parentages tested, including: J. microcarpa, (J. californica × J. nigra) × J. regia, J. hindsii × J. regia, (J. hindsii × J. regia) × J. regia, [(J. major × J. hindsii) × J. nigra] × J. regia, and J. nigra × J. regia, responses varied, but tended to be intermediate. When ‘Serr’ scions were budded or grafted on J. microcarpa × J. regia clone ‘RX1’ or Paradox (J. hindsii × J. regia) seedling rootstocks in a commercial orchard infested with P. cinnamomi, all trees on ‘RX1’ remained healthy, whereas only 49% of those on Paradox survived. Thus, useful resistance to Phytophthora is available among J. microcarpa × J. regia hybrids and is evident in ‘RX1’ rootstock.