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Root growth is critical for the effective exploitation of the rhizosphere and productive plant growth. Our recent work(1) showed that root architecture was dependent upon the degree of symplastic connectivity between neighboring cells during the specification of lateral root primordia and was affected by genes regulating callose deposition at plasmodesmata (PD). Here we provide additional evidence that both symplastic connectivity and callose are also important during the later phase of lateral root development: emergence. Callose immunolocalization assays indicated that transient symplastic isolation of the primordium occur immediately prior to emergence through the overlaying tissues to produce the mature lateral root.(1) Here we could corroborate these results by analyzing the mobility of a symplastic tracer and the expression of PD genes in lateral roots and in response to auxins. Moreover, we show that altering callose deposition affects the number of emerged lateral roots suggesting that PD regulation is important for emergence.